Stage-specific localization of cytoskeletal actin mRNA in murine seminiferous tubules and intestinal epithelia as demonstrated by in-situ hybridization

1989 ◽  
Vol 258 (2) ◽  
Author(s):  
Shigeru Sakiyama ◽  
Yohko Nakamura ◽  
Katsuo Tokunaga ◽  
Hiroshi Takazawa ◽  
Yoshinori Ohwaki ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Seminiferous Tubules ◽  
Intestinal Epithelia ◽  
Specific Localization ◽  
Actin Mrna

scholarly journals Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

1994 ◽  
Vol 42 (9) ◽  
pp. 1271-1276 ◽  
Author(s):  
M Numata ◽  
T Ono ◽  
S Iseki
Keyword(s):  
In Situ Hybridization ◽  
Significant Role ◽  
Meiotic Prophase ◽  
Oligonucleotide Probe ◽  
Mouse Testis ◽  
Seminiferous Tubules ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

scholarly journals Simultaneous in situ detection of two mRNAs in the same cell using riboprobes labeled with biotin and 35S.

1990 ◽  
Vol 38 (7) ◽  
pp. 917-922 ◽  
Author(s):  
S Ozden ◽  
C Aubert ◽  
D Gonzalez-Dunia ◽  
M Brahic
Keyword(s):  
In Situ Hybridization ◽  
Theiler's Virus ◽  
Bhk Cells ◽  
Beta Actin ◽  
Actin Mrna ◽  
High Level ◽  
Radiolabeled Probe ◽  
In Situ Detection

We used 35S-labeled and biotinylated cRNAs (riboprobes) to detect simultaneously two different mRNAs by in situ hybridization. In a first step we established the conditions under which each type of probe achieved the same high level of sensitivity. We then used these conditions to hybridize BHK cells infected with Theiler's virus, a murine picornavirus, with a mixture of a virus-specific biotinylated riboprobe and a 35S-labeled riboprobe specific for beta-actin mRNA. Both mRNAs could be detected in the same cell, although the sensitivity achieved by the radiolabeled probe was reduced by about 40% by the simultaneous hybridization with the biotinylated probe.

scholarly journals Immunohistochemistry and in situ hybridization of P-45017 alpha (17 alpha-hydroxylase/17,20-lyase).

1992 ◽  
Vol 40 (7) ◽  
pp. 903-908 ◽  
Author(s):  
T Suzuki ◽  
H Sasano ◽  
T Sawai ◽  
J I Mason ◽  
H Nagura
Keyword(s):  
In Situ Hybridization ◽  
Leydig Cells ◽  
Cellular Localization ◽  
Cdna Probe ◽  
Simultaneous Detection ◽  
Seminiferous Tubules ◽  
Zona Fasciculata ◽  
Mrna Levels ◽  
Theca Interna

Cytochrome P-45017 alpha catalyzes both 17 alpha-hydroxylation and 17,20-side-chain cleavage in steroidogenesis and lies at a key branch point in the pathways of steroid hormone biosynthesis. To obtain information on the precise localization of P-45017 alpha in swine testis, ovary, and adrenal, we undertook the simultaneous detection of P-45017 alpha mRNA and protein by combining immunohistochemistry with in situ hybridization. In situ hybridization was performed on 4% paraformaldehyde-fixed, paraffin-embedded sections by employing either a 39-base oligomer or a cDNA insert (1.7 KB) of porcine testis P-45017 alpha as DNA probe. Immunohistochemical study was performed by employing anti-P-45017 alpha. Hybridization signals were obtained in Leydig cells of the testis, theca interna of the ovarian follicle, and zona fasciculata reticularis cells of the adrenal cortex. Oligonucleotide probing yielded lower background signal than the cDNA probe. No specific signals were obtained in seminiferous tubules of the testis, medulla, and zona glomerulosa of the adrenal, and in membrana granulosa and interstitial cells of the ovary. Hybridization signals were obtained in the cells where immunoreactivity of the enzyme was observed by immunohistochemistry, except for some Leydig cells of the testis and theca interna cells of the ovary in which only immunoreactivity but not hybridization signal was observed. The present study provided detailed information about the precise cellular localization of P-45017 alpha expression at both the protein and mRNA levels in swine adrenal glands and gonads. This approach of simultaneous immunohistochemistry and in situ hybridization analysis of steroidogenic enzymes can be applied in the future to tissues exhibiting abnormal steroid metabolism and should contribute to a better understanding of steroidogenesis.

Differentiation of triticale cultivars through FISH karyotyping of their rye chromosomes

Genome ◽  
2013 ◽  
Vol 56 (5) ◽  
pp. 267-272 ◽  
Author(s):  
Maia Fradkin ◽  
María Rosa Ferrari ◽  
Shirley Mary Espert ◽  
Víctor Ferreira ◽  
Ezequiel Grassi ◽  
...  
Keyword(s):  
Cluster Analysis ◽  
In Situ Hybridization ◽  
Dna Sequences ◽  
Fish Analysis ◽  
Repetitive Dna Sequences ◽  
Agronomic Characters ◽  
Breeding Programs ◽  
Specific Localization ◽  
Highly Repetitive Dna

The aim of this work was to cytogenetically characterize triticale cultivars through fluorescence in situ hybridization (FISH) analysis of their rye chromosomes. In the present work, we studied six cultivars of triticale (‘Cayú-UNRC’, ‘Cumé-UNRC’, ‘Genú-UNRC’, ‘Ñinca-UNRC’, ‘Quiñé-UNRC’, and ‘Tizné-UNRC’), released by the Universidad Nacional de Río Cuarto (UNRC), Córdoba, Argentina. The cultivars were obtained from the International Center for the Improvement of Maize and Wheat (CIMMYT) and improved for fresh forage, haymaking, and feed grain at UNRC. The distribution and organization of highly repetitive DNA sequences of Secale cereale (pSc74, pSc200, pSc250, and pSc119.2) using FISH analyses revealed a specific localization of the signals for several rye chromosomes, which allowed us to distinguish the cultivars. Cluster analysis showed a great cytogenetic similarity among the rye cultivars used to originate these hybrids. The knowledge of the variability among triticale cultivars is necessary to propose future crosses in breeding programs. This study will also be valuable to identify commercial seeds and to analyze the possible association between agronomic characters and the presence of certain rye chromosomes or specific regions in these chromosomes.

scholarly journals The localization of actin mRNA in megakaryocytes demonstrated by in situ hybridization using digoxigenin-labeled riboprobes.

1991 ◽  
Vol 24 (3) ◽  
pp. 335-339
Author(s):  
JUNZO SASAKI ◽  
TAKAKO NOMURA ◽  
SADAHIRO WATANABE ◽  
SHIGETO KANDA ◽  
TOSHIYUKI KOUNO ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Actin Mrna

scholarly journals Localization of protamine 1 mRNA in different stages of the cycle of the rat seminiferous epithelium.

1988 ◽  
Vol 107 (2) ◽  
pp. 407-412 ◽  
Author(s):  
P Mali ◽  
M Sandberg ◽  
E Vuorio ◽  
P C Yelick ◽  
N B Hecht ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Northern Blot ◽  
Northern Blot Analysis ◽  
Cdna Probe ◽  
Seminiferous Epithelium ◽  
Seminiferous Tubules ◽  
Mrna Levels ◽  
Low Levels ◽  
Protamine 1

A mouse protamine 1 cDNA probe was used to study P1 protamine gene expression during the cycle of the seminiferous epithelium in the rat. In situ hybridization experiments showed that transcription of the P1 protamine mRNA starts in the middle of step 7 of spermiogenesis during substage VIIc. The mRNA levels stay high in steps 7-14 spermatids but decrease during steps 15-16 and are virtually undetectable in steps 17-19 spermatids. Northern blot analyses of RNAs isolated from microdissected pools of seminiferous tubules show high P1 protamine mRNA concentrations during stages VIIc-XIV-III of the cycle and lower levels during stages IV-VIIb. Owing to a post-transcriptional shortening of the poly(A) tail by 130 bases, a decrease in the size of protamine 1 mRNA from approximately 580 to 450 nucleotides was observed in stages XIII-XIV suggesting an initiation of protamine 1 synthesis in step 13-14 spermatids. In stages II-VI (steps 16-18 spermatids), only the smaller size protamine 1 mRNA was detectable. The expression of protamine 1 mRNAs has been localized in the very last phase of the haploid gene activity. Although the in situ hybridization suggests a disappearance of protamine 1 mRNA after step 16 of spermiogenesis, Northern blot analysis shows that low levels of mRNA are present during the period of final condensation of the chromatin, reflecting the association of protamine with DNA.

scholarly journals Period gene expression in mouse endocrine tissues

2003 ◽  
Vol 285 (3) ◽  
pp. R561-R569 ◽  
Author(s):  
Eric L. Bittman ◽  
Leo Doherty ◽  
Liyue Huang ◽  
Allison Paroskie
Keyword(s):  
In Situ Hybridization ◽  
Cell Types ◽  
Seminiferous Tubules ◽  
Double Labeling ◽  
Peripheral Tissues ◽  
Period Gene ◽  
Central Brain ◽  
Tissue Compartments ◽  
Endocrine Tissues

Circadian rhythms are generated by the oscillating expression of the Per1 and Per2 genes, which are expressed not only in the central brain pacemaker but also in peripheral tissues. Hormones are likely to coordinate physiological function in time. We performed in situ hybridization to localize mPer1 and mPer2 mRNA to particular cell types and tissue compartments in adrenal, thyroid, and testis. BALB/c mice maintained in a 12:12-h light-dark cycle expressed mPer1 in adrenal medulla, particularly in late afternoon and early night. mPer2 mRNA was more intensely expressed in adrenal cortex, especially in afternoon and evening. mPer1 mRNA was detected in thyroid. mPer1 was found in some but not all seminiferous tubules of each mouse at all times of day. Quantitation in C57BL/6 mice revealed a significant increase in the number of heavily labeled seminiferous tubules early in the night. Consistent with in situ hybridization, immunocytochemistry showed PER1 protein in spermatocytes and spermatids (spermatogenic stages VII-XII). Staining in spermatogonia and interstitial cells was inconsistent. Double labeling with 5′-bromodeoxyuridine showed PER1 expression first occurring 5 days after DNA replication. We conclude that mPeriod genes are expressed in peripheral endocrine glands. Central regulation, adenohypophyseal control, and functional importance of expression and phase remain to be elucidated.

scholarly journals Tissue-Specific Localization of Cytochrome P450 Aromatase in the Equine Embryo by In Situ Hybridization and Immunocytochemistry1

2000 ◽  
Vol 62 (5) ◽  
pp. 1141-1145 ◽  
Author(s):  
Karen W. Walters ◽  
C. Jo Corbin ◽  
Gary B. Anderson ◽  
Janet F. Roser ◽  
Alan J. Conley
Keyword(s):  
Cytochrome P450 ◽  
In Situ Hybridization ◽  
Tissue Specific ◽  
P450 Aromatase ◽  
Cytochrome P450 Aromatase ◽  
Specific Localization

scholarly journals Visualization of intestinal infections with astro- and sapovirus in mink (Neovison vison) kits by in situ hybridization

FEMS Microbes ◽  
2021 ◽  
Author(s):  
Julie Melsted Birch ◽  
Mikael Leijon ◽  
Søren Saxmose Nielsen ◽  
Tina Struve ◽  
Henrik Elvang Jensen
Keyword(s):  
In Situ Hybridization ◽  
Goblet Cells ◽  
Neovison Vison ◽  
Localization Pattern ◽  
Intestinal Infections ◽  
Specific Localization ◽  
The Impact ◽  
First Time ◽  
Mink Kits

Abstract Clarification of the infection microbiology remains a challenge in the pre-weaning diarrhea (PWD) syndrome in farmed mink (Neovison vison). Duodenal, jejunal and colon sections from 36 mink kits with PWD were systematically examined by chromogen in situ hybridization (CISH) targeting two incriminated viruses: Mink astrovirus and mink sapovirus. Using the RNAscope® 2.5 HD Duplex Assay astrovirus and sapovirus were visualized and simultaneously demonstrated in the gut tissue. Both viruses infect enterocytes in the small intestine with a specific localization pattern; astrovirus affects the two apical thirds of the villi, whereas sapovirus generally affects the basal parts of the villi. Furthermore, we demonstrated that astrovirus in mink does not target the goblet cells. This is the first time astro- and calicivirus have been visualized in mink kit gut tissue, and these findings might be important in clarification of the impact of these viruses in the PWD syndrome.

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