Infertility due to defective sperm flagella caused by an intronic deletion in DNAH17 that perturbs splicing

Abstract Artificial insemination in pig (Sus scrofa domesticus) breeding involves the evaluation of the semen quality of breeding boars. Ejaculates that fulfill predefined quality requirements are processed, diluted and used for inseminations. Within short time, eight Swiss Large White boars producing immotile sperm that had multiple morphological abnormalities of the sperm flagella were noticed at a semen collection center. The eight boars were inbred on a common ancestor suggesting that the novel sperm flagella defect is a recessive trait. Transmission electron microscopy cross-sections revealed that the immotile sperm had disorganized flagellar axonemes. Haplotype-based association testing involving microarray-derived genotypes at 41,094 SNPs of six affected and 100 fertile boars yielded strong association (P = 4.22 × 10−15) at chromosome 12. Autozygosity mapping enabled us to pinpoint the causal mutation on a 1.11 Mb haplotype located between 3,473,632 and 4,587,759 bp. The haplotype carries an intronic 13-bp deletion (Chr12:3,556,401–3,556,414 bp) that is compatible with recessive inheritance. The 13-bp deletion excises the polypyrimidine tract upstream exon 56 of DNAH17 (XM_021066525.1: c.8510–17_8510–5del) encoding dynein axonemal heavy chain 17. Transcriptome analysis of the testis of two affected boars revealed that the loss of the polypyrimidine tract causes exon skipping which results in the in-frame loss of 89 amino acids from DNAH17. Disruption of DNAH17 impairs the assembly of the flagellar axoneme and manifests in multiple morphological abnormalities of the sperm flagella. Direct gene testing may now be implemented to monitor the defective allele in the Swiss Large White population and prevent the frequent manifestation of a sterilizing sperm tail disorder in breeding boars.

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Infertility due to defective sperm flagella caused by an intronic deletion in DNAH17 that perturbs splicing

10.1101/2020.10.16.333344 ◽

2020 ◽

Author(s):

Adéla Nosková ◽

Maya Hiltpold ◽

Fredi Janett ◽

Thomas Echtermann ◽

Zih-Hua Fang ◽

...

Keyword(s):

Cross Sections ◽

Semen Quality ◽

Strong Association ◽

Exon Skipping ◽

Recessive Trait ◽

Polypyrimidine Tract ◽

Large White ◽

Morphological Abnormalities ◽

Association Testing ◽

Gene Testing

ABSTRACTArtificial insemination in pig (Sus scrofa domesticus) breeding involves the evaluation of the semen quality of breeding boars. Ejaculates that fulfill predefined quality requirements are processed, diluted and used for inseminations. Within short time, eight Swiss Large White boars producing immotile sperm that had multiple morphological abnormalities of the sperm flagella were noticed at a semen collection center. The eight boars were inbred on a common ancestor suggesting that the novel sperm flagella defect is a recessive trait. Transmission electron microscopy cross-sections revealed that the immotile sperm had disorganized flagellar axonemes. Haplotype-based association testing involving microarray-derived genotypes at 41,094 SNPs of six affected and 100 fertile boars yielded strong association (P=4.22 × 10−15) at chromosome 12. Autozygosity mapping enabled us to pinpoint the causal mutation on a 1.11 Mb haplotype located between 3,473,632 and 4,587,759 bp. The haplotype carries an intronic 13-bp deletion (Chr12:3,556,401-3,556,414 bp) that is compatible with recessive inheritance. The 13-bp deletion excises the polypyrimidine tract upstream exon 56 of DNAH17 (XM_021066525.1:c.8510-17_8510-5del) encoding dynein axonemal heavy chain 17. Transcriptome analysis of the testis of two affected boars revealed that the loss of the polypyrimidine tract causes exon skipping which results in the in-frame loss of 89 amino acids from DNAH17. Disruption of DNAH17 impairs the assembly of the flagellar axoneme and manifests in multiple morphological abnormalities of the sperm flagella. Direct gene testing may now be implemented to monitor the defective allele in the Swiss Large White population and prevent the frequent manifestation of a sterilizing sperm tail disorder in breeding boars.

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Semen quality of three breeds of exotic boar extended using methaloninated Moringa leaf extract as antibiotics in beltsville thawing solution

Nigerian Journal of Animal Production ◽

10.51791/njap.v48i1.2897 ◽

2021 ◽

Vol 48 (1) ◽

pp. 47-54

Author(s):

I. A. Omodewu ◽

T. Olorungbohunmi ◽

B. A. Boladuro ◽

J. O. Abiona ◽

C. P. Njoku

Keyword(s):

Leaf Extract ◽

Storage Time ◽

Moringa Oleifera ◽

Semen Quality ◽

Antibacterial Properties ◽

Semen Parameters ◽

Large White ◽

Normal Limits ◽

Qualité Du Sperme

Moringa oleifera leaf extract (MLE) contains antibacterial properties; this study evaluated the effect of including MLE, breed and storage time on semen quality of boars extended with Beltsville thawing solution (BTS). Semen samples were collected weekly from twelve boars (Four Duroc, four Large White and four Landrace). Samples were extended with 0, 0.25, 0.50, 0.75 and 1.00g MLE inclusions. Parameters; volume and concentration were taken immediately after collection while pH, mass activity (%MA), progressive motility (%PM), liveability (%LA) and abnormality (%ABN) were taken at 0, 24, 48 and 72 hours. Data obtained were subjected to analysis of variance. All parameters considered except %ABN were significant (p<0.05). MA ranged between 3.31 (1.00gMLE) and 4.00 (0gMLE). Highest PM was 60.93% in 0.25gMLE, pH differed with 6.98 (0gMLE) and 6.90 (1.00gMLE), 0gMLE (89.64%) had higher LA compared to 0.25gMLE (86.40%), ABN ranged between3.94 to 4.60%. As storage time increased viability of all semen parameters reduced, between 48 and 72 hours parameters reduced below normal limits. Breed effect differed (p<0.05) for MA, Volume and Conc., Duroc (3.78) had higher MA compared to Large White (3.64) and Landrace (3.60). Semen volume (p<0.05) for Landrace (255.20g) was highest while Duroc (170.80g) was lowest. Large white concentration (201.8million spermatozoa/ml) was higher (p<0.05) than Landrace (187.4 million spermatozoa/ml) and Duroc (123.8million spermatozoa/ml). %LA ranged between 88.06% and 88.72% for Duroc and landrace respectively while %ABN was between 3.97-4.41% for Large White and Landrace. This study concluded that 0.75-1.00gMLE could replace synthetic antibiotic in BTS for 24 hours. L'extrait de feuille de Moringa oleifera (le 'MLE') contient des propriétés antibactériennes ; Cette étude a évalué l'effet de l'inclusion de 'MLE', de la race et du temps de stockage sur la qualité du sperme de verrats exotiques additionnés de solution de décongélation de Beltsville (le 'BTS'). Des échantillons de sperme ont été prises chaque semaine de douze verrats (quatre Duroc, quatre Large White et quatre Landrace). Les échantillons ont été étendus avec des inclusions de 0, 0,25, 0,50, 0,75 et 1,00 g de 'MLE'. Paramètres ; le volume et la concentration ont été prélevés immédiatement après le prélèvement tandis que le pH, l'activité massique (% 'MA'), la motilité progressive (% 'PM'), l'habitabilité (% LA) et l'anomalie (% ABN) ont été pris à 0, 24, 48 et 72 heures. Les données obtenues ont été soumises à une analyse de variance. Tous les paramètres considérés à l'exception du% ABN étaient significatifs (p <0,05). Le MA variait entre 3,31 (1,00 g MLE) et 4,00 (0 gMLE). La'PM la plus élevée était de 60,93% dans 0,25 gMLE, le pH différait de 6,98 (0 gMLE) et 6,90 (1,00 gMLE), 0 gMLE (89,64%) avait un LA plus élevé par rapport à 0,25 gMLE (86,40%), l''ABN' variait entre 3,94 et 4,60%. À mesure que la durée de stockage augmentait la viabilité de tous les paramètres du sperme, entre 48 et 72 heures, les paramètres étaient réduits en dessous des limites normales. L'effet de la race était différent (p <0,05) pour la MA, le volume et la concentration. Duroc (3,78) avait une MA plus élevée que le Large White (3,64) et le Landrace (3,60). Le volume de sperme (p <0,05) pour Landrace (255,20 g) était le plus élevé, tandis que Duroc (170,80 g) était le plus faible. La grande concentration de blanc (201,8 millions de spermatozoïdes / ml) était plus élevée (p <0,05) que Landrace (187,4 millions de spermatozoïdes / ml) et Duroc (123,8 millions de spermatozoïdes / ml). % LA variait entre 88,06% et 88,72% pour Duroc et landrace respectivement tandis que% ABN était entre 3,97-4,41% pour Large White et Landrace. Cette étude a conclu que 0,75-1,00 gMLE pouvait remplacer l'antibiotique synthétique dans le BTS pendant 24 heures.

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111 ESTIMATION OF SPERM QUALITY IN FRESH AND FROZEN - THAWED SEMEN FROM ASTURIANA DE LOS VALLES BULLS

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab111 ◽

2006 ◽

Vol 18 (2) ◽

pp. 163

Author(s):

C. Tamargo ◽

M. Carbajo ◽

C. Diez ◽

D. Martin ◽

C. O. Hidalgo

Keyword(s):

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Daily Routine ◽

Morphological Abnormalities ◽

Hypoosmotic Swelling Test ◽

Motion Characteristics ◽

The Impact ◽

Casa System

Artificial insemination and semen cryopreservation have significantly improved the breeding potential of male animals. However, current freezing techniques commonly result in reduced semen quality (Januskauskas et al. 1999 Theriogenology 52, 641-658), and surviving cells are affected post-thaw either structurally or functionally (Nagy et al. 2004 Anim. Reprod. Sci. 80, 225-235). In this work we analyze the impact of cryopreservation on Asturiana de los Valles bull sperm. Ejaculates (n = 373) from seven adult bulls were weekly collected by means of artificial vagina. Immediately after collection, routine parameters including volume (V), mass motility (MM), and concentration (C) of sperm cells were evaluated. Then the semen was extended with a commercial extender, loaded into 0.25-mL plastic straws at a concentration of 23 � 106 per straw, frozen and stored for further analysis. Four straws per ejaculate were thawed, pooled and analyzed for motion characteristics by means of a CASA system (Sperm Class Analyzer, SCA 2002� Microptic S. L., Barcelona, Spain) added to an optical phase-contrast microscope with heatable (37�C) stage. Immediately after thawing, we analyzed the % of motile spermatozoa (MS) and the % of progressively motile spermatozoa (PMS); then samples were incubated for 3 h at 37�C and MS and PMS were measured again (MS3 and PMS3, respectively). Functional integrity of the plasmallema was evaluated by the hypoosmotic swelling test (HOST) together with the % of typical tail coiling/swelling (percentage of HOST-positive spermatozoa, HOST-PS). The % of viable spermatozoa (VS) [membrane integrity was evaluated by fluorescence microscopy with a dual staining system (propidium iodide (PI) and 6-carboxyfluorescein diacetate (CFDA)]. Sperm showing partial or complete red fluorescence (PI staining) were considered nonviable, whereas sperm showing complete green fluorescence were considered viable. Altered acrosomes (AA) and morphological abnormalities were also determined. The % of morphological abnormalities was classified according to their location in head (HA), midpiece (MA), and tail (TA). Proximal and distal cytoplasmic droplets were counted as separate abnormalities (CD). Data were analyzed by the MEANS procedure of SAS (SAS Institute, Inc., Cary, NC, USA). A significant (P < 0.05) decrease in the sperm motility was observed after freezing/thawing (MS: 80.20 � 0.75 vs. 47.36 � 1.04, and PMS: 68.73 � 0.73 vs. 42.14 � 0.96 for fresh and frozen-thawed semen, respectively). Also, the frozen-thawed sperm showed increased % of HA, MA, AA, HOST-PS, and VS (P < 0.05). These morphological abnormalities could contribute to decreasing sperm motility. The new computer and video technologies provide useful information about sperm quality and can be used in the daily routine of processing semen. This work was performed in collaboration with ASEAVA.

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A 1-bp deletion in bovine QRICH2 causes low sperm count and immotile sperm with multiple morphological abnormalities

10.1101/2021.11.19.469233 ◽

2021 ◽

Author(s):

Maya Hiltpold ◽

Fredi Janett ◽

Xena Marie Mapel ◽

Naveen Kumar Kadri ◽

Zih-Hua Fang ◽

...

Keyword(s):

Premature Termination ◽

Semen Quality ◽

Sperm Count ◽

Sperm Concentration ◽

Premature Termination Codon ◽

Termination Codon ◽

Sequence Variant ◽

Loss Of Function ◽

Morphological Abnormalities ◽

Brown Swiss

Background: Semen quality and male fertility are monitored in artificial insemination bulls to ensure high insemination success rates. Only ejaculates that fulfill minimum quality requirements are processed and eventually used for artificial inseminations. We examined 70,990 ejaculates from 1343 Brown Swiss bulls to identify bulls from which all ejaculates were rejected due to low semen quality. This procedure identified a bull that produced twelve ejaculates with an aberrantly low number of sperm (0.2±0.2 x 109 sperm per ml) which were mostly immotile due to multiple morphological abnormalities. Results: The genome of the bull was sequenced at 12-fold coverage to investigate a suspected genetic cause. Comparing the sequence variant genotypes of the bull with those from 397 fertile bulls revealed a 1-bp deletion in the coding sequence of QRICH2 encoding glutamine rich 2 as a compelling candidate causal variant. The 1-bp deletion causes a frameshift in translation and induces a premature termination codon (ENSBTAP00000018337.1:p.Cys1644AlafsTer52). The analysis of testis transcriptomes from 76 bulls showed that the transcript with the premature termination codon is subjected to nonsense-mediated mRNA decay. The 1-bp deletion resides on a 675 kb haplotype spanning 181 SNPs from the Illumina BovineHD Bead chip. The haplotype segregates at a frequency of 5% in the Brown Swiss cattle population. This analysis also identified another bull that carried the 1-bp deletion in the homozygous state. Semen analyses from the second bull confirmed low sperm concentration and immotile sperm with multiple morphological abnormalities primarily affecting the sperm flagellum and, to a lesser extent, the sperm head. Conclusions: A recessive loss-of-function allele of bovine QRICH2 likely causes low sperm concentration and immotile sperm with multiple morphological abnormalities. Routine sperm analyses unambiguously identify homozygous bulls. A direct gene test can be implemented to monitor the frequency of the undesired allele in cattle populations.

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Cooperation of pre-mRNA sequence elements in splice site selection

Molecular and Cellular Biology ◽

10.1128/mcb.12.5.2108-2114.1992 ◽

1992 ◽

Vol 12 (5) ◽

pp. 2108-2114

Author(s):

Z Dominski ◽

R Kole

Keyword(s):

Splice Site ◽

Hela Cells ◽

Site Selection ◽

Exon Skipping ◽

Polypyrimidine Tract ◽

Splice Site Selection ◽

Branch Point Sequence ◽

Sequence Elements

We have recently demonstrated that short internal exons in pre-mRNA transcripts with three exons and two introns are ignored by splicing machinery in vitro and in vivo, resulting in exon skipping. Exon skipping is reversed when the pyrimidine content of the polypyrimidine tract in the upstream intron is increased (Z. Dominski and R. Kole, Mol. Cell. Biol. 11:6075-6083, 1991). Here we show that skipping of the short internal exon can be partially reversed by mutations which modify the upstream branch point sequence of the 5' splice site at the end of the exon to their respective consensus sequences. When the modified elements are combined with one another in the same pre-mRNA, exon skipping is fully reversed. Full reversion of exon skipping is also observed when these elements are combined individually with the upstream polypyrimidine tract strengthened by three purine-to-pyrimidine mutations. The observed patterns of splice site selection are similar in vitro (in nuclear extracts from HeLa cells) and in vivo (in transfected HeLa cells). We also show that the length of the downstream intron plays a role in splice site selection. Our data indicate that the interplay between the sequence elements in pre-mRNA controls the outcome of each splicing event, providing the means for very subtle regulation of alternative splicing.

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Boar sperm quality in relation to presence of sp32-like protein in spermatozoa - preliminary studies

Bulletin of the Veterinary Institute in Pulawy ◽

10.1515/bvip-2015-0041 ◽

2015 ◽

Vol 59 (2) ◽

pp. 279-286 ◽

Cited By ~ 1

Author(s):

Aleksandra Orzołek ◽

Paweł Wysocki ◽

Jerzy Strzeżek ◽

Magdalena Koziorowska-Gilun ◽

Anna Dziekońska ◽

...

Keyword(s):

Sperm Motility ◽

Seminal Plasma ◽

Semen Quality ◽

Sperm Quality ◽

Total Antioxidant Status ◽

Atp Content ◽

Large White ◽

Sod Activity ◽

Spermatozoa Motility ◽

Total Antioxidant

Abstract The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW) and Polish Landrace (PL) boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility, lipid peroxidation levels (MDA production), ATP content, activities of superoxide dismutase (SOD) and catalase (CAT), total antioxidant status (TAS), and total oxidant status (TOS) of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32). The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility, lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality

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20 SPERM CHARACTERIZATION OF ASTURCON PONIES AFTER COLLECTION AND EQUILIBRATION/REFRIGERATION BEFORE FREEZING

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab20 ◽

2007 ◽

Vol 19 (1) ◽

pp. 128

Author(s):

C. Tamargo ◽

C. Díez ◽

J. De La Fuente ◽

M. Carbajo ◽

J. M. Benito ◽

...

Keyword(s):

Biological Diversity ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Skim Milk ◽

Egg Yolk ◽

Morphological Abnormalities ◽

Unique Source ◽

And Performance ◽

Breeding Soundness

The need to conserve farm animal biodiversity is accepted by many countries through the ratification of the convention of biological diversity, and sperm quality is known to be an important criterion in the evaluation of breeding soundness. The aim of this work was to characterize the semen of a local breed of ponies 'Asturcon' (maintained free over the mountains all year around) before its incorporation into a germplasm bank. Semen was obtained from six stallions (6–17 years of age) using an artificial vagina, 3 days/week, during 12 weeks. Immediately after collection, gel-free semen was evaluated for volume, sperm concentration, and motility. Semen motility was again evaluated after equilibration/refrigeration. For evaluation of individual (IM) and progressive motility (PM) rates, semen was diluted (20 � 106 spermatozoa/mL) and analyzed with a CASA System (SCA; Microptic S.L., Barcelona, Spain). Five fields per sample were evaluated (minimum 500 spermatozoa/sample) under a phase contrast microscope (100�). Semen samples were subjected to a hypo-osmotic swelling test (HOS) test to detect the presence of swollen tails in a 100 mM citrate–fructose solution. Percentages of altered acrosomes and morphological abnormalities were determined by counting 100 spermatozoa (1000�). Then, semen was diluted and centrifuged for 10 min at 600g. After the supernatant was discarded, the pellet was re-suspended in freezing medium (skim milk extender containing 2% egg yolk and 2.5% glycerol) to a final concentration of 100 � 106 spermatozoa/mL, and equilibrated/cooled (60 min) to 4�C. Statistical analysis was carried out by means of the GLM and CORR procedures and Duncan test for means (SAS Institute, Inc., Cary, NC, USA). A significant effect between males (P < 0.05) on semen quality, such as volume of the ejaculate, sperm concentration, and morphological abnormalities, were detected among stallions. On the other hand, positive and significant correlations were found between the sperm motility immediately after collection and after equilbration/refrigeration (r = 0.73; P < 0.05); moreover, sperm motilities (both fresh and refrigerated) correlated with the results of the HOS test (r = 0.56; P < 0.001, and r = 0.27, P < 0.05, respectively). These preliminary results confirm that the sperm of the Asturcon ponies breed can be collected and will survive the equilibration/refrigeration procedures. Conservation and development of local breeds is important because they represent a unique source of genes for improving health and performance traits of industrial breeds. However, complementary studies on the ability of the stallion sperm to survive freezing/thawing procedures in rates higher than 30% are needed to ensure that genetic banks are correctly created. This work was performed in collaboration with ACPRA and Dep�sito de Sementales de Santander (Spain), and supported by RZ2004-00031-C02-01.

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21 Comparative Study on Epididymal Spermatozoa Traits of Large White×Landrace and Kolbroek Boars

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab21 ◽

2018 ◽

Vol 30 (1) ◽

pp. 150

Author(s):

T. R. Netshirovha ◽

A. T. Kanengoni ◽

M. B. Matabane ◽

M. L. Mphaphathi ◽

A. Maqhashu ◽

...

Keyword(s):

Semen Quality ◽

Large White ◽

Sperm Analysis ◽

Spermatozoa Motility ◽

Average Percentage ◽

Semen Volume ◽

Significant Difference ◽

Curve Line ◽

Epididymal Spermatozoa ◽

The Difference

The Kolbroek pig is an early maturing breed (maturing at 4 to 5 months) that grows more slowly than modern pig breeds. There is general concern that the genetic variation within Kolbroek pig breed is becoming extinct. The aim of the study was to compare semen quality of epididymal spermatozoa derived from slaughtered Large White × Landrace and Kolbroek boars aged between 7 and 9 months. Kolbroek (n = 10) and Large White × Landrace (n = 10) boars were used in this study. Semen was collected from the head of epididymis of Kolbroek and Large White × Landrace testicles. Spermatozoa samples were extracted from head of epididymis by making incision with a razor. Semen samples were then evaluated for macroscopic (semen volume, pH, and concentration) and microscopic characteristics (spermatozoa motility and morphology). Spermatozoa motility was evaluated using computer-aided sperm analysis. Analysis of variance was used to test the difference between the breeds. The average percentage of Kolbroek and Large White × Landrace spermatozoa total motility was 92.4 ± 4.0 and 94.0 ± 4.1%, respectively (P > 0.05). The spermatozoa velocity on the curve line for Kolbroek was lower (147.2 ± 39.2) compared with the Large White × Landrace (178.3 ± 30.1 μm/s; P > 0.05). However, no significant difference was observed between the 2 breeds for rapid spermatozoa and total spermatozoa motility. No significant differences were observed in Kolbroek and Large White × Landrace boar semen volume (8 and 9 mL, respectively) or semen pH (7.0). The average spermatozoa concentration for Kolbroek and Large White × Landrace was 2.5 ± 1.2 and 1.1 ± 1.0 (×109 mL−1), respectively (P > 0.05). There were no significant differences between Kolbroek and Large White × Landrace in abnormal spermatozoa morphology. However, spermatozoa with distal, head, and midpiece abnormalities were significantly different in Kolbroek (14.0 ± 3.6; 3.6 ± 6.0 and 3.4 ± 3.7%) and Large White × Landrace breed (5.4 ± 4.7; 4.7 ± 1.0 and 0.4 ± 1.0%), respectively. In conclusion, macroscopic and microscopic epididymal spermatozoa characteristics of Kolbroek were similar to those of Large White × Landrace boars, except for head and distal spermatozoa abnormalities.

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Cooperation of pre-mRNA sequence elements in splice site selection.

Molecular and Cellular Biology ◽

10.1128/mcb.12.5.2108 ◽

1992 ◽

Vol 12 (5) ◽

pp. 2108-2114 ◽

Cited By ~ 31

Author(s):

Z Dominski ◽

R Kole

Keyword(s):

Splice Site ◽

Hela Cells ◽

Site Selection ◽

Exon Skipping ◽

Polypyrimidine Tract ◽

Splice Site Selection ◽

Branch Point Sequence ◽

Sequence Elements

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Detection of one large insertion/deletion (indel) and two novel SNPs within the <i>SPEF2</i> gene and their associations with male piglet reproduction traits

Archives Animal Breeding ◽

10.5194/aab-59-275-2016 ◽

2016 ◽

Vol 59 (2) ◽

pp. 275-283 ◽

Cited By ~ 12

Author(s):

Rui Chen ◽

Shuai Yu ◽

Fa Ren ◽

Xiao Yan Lv ◽

Chuan Ying Pan

Keyword(s):

Dna Marker ◽

Male Reproduction ◽

Nucleotide Polymorphisms ◽

Chromosome 16 ◽

Large White ◽

Single Nucleotide ◽

Association Testing ◽

Normal Sperm ◽

Reproduction Traits ◽

Large Insertion

Abstract. The sperm flagella 2 (SPEF2) gene is essential for normal sperm tail development and male fertility. To fully characterize the structure of the mutation and to further study the function of the pig SPEF2 gene, we explored the insertion/deletion (indel) and novel single-nucleotide polymorphisms (SNPs) within the pig SPEF2 gene, and tested their associations with the testicular sizes in male Large White (LW) and Landrace (LD) pigs from China. Herein, a large insertion located at the SPEF2 gene in chromosome 16 was found, and two alleles of "I" (insertion) and "D" (deletion) were designated. Allele "D" was dominant in all analyzed pigs. Two novel SNPs (namely (NC_010458) g.19642G > A, resulting in AfaI aCRS PCR–PFLP, and g.19886C > G, resulting in EcoRI aCRS PCR–PFLP) were found in LW and LD pigs. Association testing revealed that g.19886C > G was significantly associated with the testis long circumference (TLC) in LW pigs (P < 0.05), suggesting that this SNP would be the DNA marker for the marker-assisted selection (MAS) in reproduction traits. This preliminary result indicates that the pig SPEF2 gene had significant effects on male reproduction traits. These findings could not only extend the spectrum of genetic variations in the pig SPEF2 gene but also contribute to implementing MAS in genetics and breeding in pigs.

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