Morphometric, Histochemical, and Ultrastructural Analysis of the Reproductive System and Spermatogenic Stages of Male Blue Crab (Callinectessapidus Rathbun, 1896)

The blue crab, Callinectes sapidus Rathbun, 1896, is considered to be a luxury meal, especially in touristic cities. It contains more than 20 types of amino acids and provides all the needed amounts for human growth. This study describes the morphology and complex differentiation in the gonadosomatic index, morphological and ultrastructure features of the reproductive system, spermatogenesis, and spermatophores structure; this is due to the need to maintain natural and fishing stocks. Mature adult male crabs (carapace length 59 ± 7.12; width 126 ± 18.8 mm) were obtained from Abu-Qir Bay from November 2018 to October 2019 and transported alive in seawater to the laboratory. The reproductive system was dissected and weighed to the nearest 0.001g using the electronic balance, and the gonadosomatic index was subsequently calculated. The morphological analysis showed the developing testes with highly compacted seminiferous tubules. Using Periodic acid–Schiff stain, the spermatophore appeared with a zigzag-shaped wall that indicates its carbohydrate constituents. Each Spermatophore consisted of an inner spermatozoal mass embedded in a matrix, whose main components were secretions 1 and 2, and an outer thin acellular layer composed of secretions 3 and 4 from the anterior vas deferens (AVD). Secretions 5 and 6 (S5 and S6) also appeared with carbohydrate constituents using Mallory triple stain. The transverse section of the middle vas deferens (MVD) showed spermatophores with rod-shaped secretion S6 and granular secretions 7 (S7), forming a complex matrix between spermatophores. The secretion found in the MVD was granular, strongly acidophilic, and secreted by its highly columnar epithelium (S7). The ultrastructure showed that the testes were surrounded by a monolayer of myoid cells with an elongated nucleus, which also contained the following stages: spermatogonia, primary spermatocyte, secondary spermatocytes, and spermatids as well as spermatozoa. On the other hand, scanning electron microscope studies for fully formed spermatophore taken from the middle part of the vas deferens indicated that it is ellipsoidal in its outline with terminal stalk. Furthermore, the spermatophore was surrounded by a thick capsule of non-cellular substances and contained mature spermatozoa.

Download Full-text

Morphological and Histological Features of the Vomeronasal Organ in African Pygmy Hedgehog (Atelerix albiventris)

Animals ◽

10.3390/ani11051462 ◽

2021 ◽

Vol 11 (5) ◽

pp. 1462

Author(s):

Daisuke Kondoh ◽

Yusuke Tanaka ◽

Yusuke K. Kawai ◽

Takayuki Mineshige ◽

Kenichi Watanabe ◽

...

Keyword(s):

Soft Tissues ◽

Periodic Acid ◽

Vomeronasal Organ ◽

Sensory Epithelium ◽

Middle Part ◽

Periodic Acid Schiff ◽

Histological Features ◽

Duct Opening ◽

Hematoxylin Eosin ◽

Rostral Part

The vomeronasal organ (VNO) detects specific chemicals such as pheromones and kairomones. Hedgehogs (Eulipotyphla: Erinaceidae) have a well-developed accessory olfactory bulb that receives projections from the VNO, but little is known about the hedgehog VNO. Here, we studied the histological features of the VNO in five individual African pygmy hedgehogs by hematoxylin-eosin, periodic acid-Schiff, and Alcian blue stains. The hedgehog VNO comprises a hyaline cartilage capsule, soft tissue and epithelial lumen, and it branches from the site just before the incisive duct opening into the nasal cavity. The soft tissues contain several small mucous (or mucoserous) glands and a large serous gland, and many venous sinuses all around the lumen. The VNO lumen is round to oval throughout the hedgehog VNO, and the sensory epithelium lines almost the entire rostral part and medial wall of the middle part. These findings indicate that the VNO is functional and plays an important role in the hedgehog. Notably, the VNO apparently has a characteristic flushing mechanism with serous secretions like those of gustatory glands, which the hedgehog might frequently use to recognize the external environment.

Download Full-text

Morphology of male reproductive organs of the rice field crab Esanthelphusa nani (Naiyanetr, 1984) (Brachyura, Gecarcinucidae)

Crustaceana ◽

10.1163/15685403-bja10130 ◽

2021 ◽

Vol 94 (7) ◽

pp. 823-844

Author(s):

Rachata Maneein ◽

Julien Claude ◽

Jirarach Kitana ◽

Noppadon Kitana

Keyword(s):

Rice Field ◽

Reproductive Tract ◽

Vas Deferens ◽

Gonadosomatic Index ◽

Relative Weight ◽

Reproductive Organs ◽

Seminiferous Tubules ◽

Adult Males ◽

Natural Habitats ◽

Tropical Regions

Abstract The rice field crab is considered a pest that disrupts rice production in paddy fields in tropical regions. The crab is also used as a stable food source by local people in many countries in Southeast Asia. For both of these roles, a good knowledge of the crab’s reproductive biology is necessary to manage populations in their natural habitats. This study aims to investigate the reproductive cycle of adult males of the rice field crab Esanthelphusa nani (Naiyanetr, 1984). Males crabs were sampled monthly in a paddy field; individuals were then anaesthetized and dissected, and their reproductive tract was analysed. The gonadosomatic index (GSI) was evaluated based on the relative weight of the reproductive tract and body weight. The male GSI showed significant temporal difference, with the highest levels in June, August and September. Based on GSI and macroscopic examination, the testis and vas deferens can be classified into three stages of increasing size: (1) resting stage, (2) developing stage and (3) active stage. Microscopic examination revealed that the testis and vas deferens had a tubular structure. Seminiferous tubules in the testis underwent spermatogenesis and spermiogenesis at every stage. The vas deferens consisted of three major regions: anterior, middle and posterior, which showed different types of epithelia. Interestingly, the microscopic structure of the vas deferens was similar among these three reproductive stages. It is also important to note that round-shaped spermatozoa were found in different parts of the reproductive tract, including the seminiferous tubule and vas deferens, as well as in the female seminal receptacle, whereas no spermatophores were found in this species. Overall, this study shows that mature sperms can be found in the reproductive tract of male E. nani throughout the year, indicating that males have prolonged spermatogenic activities.

Download Full-text

Acute Exposure to the Neonicotinoid Insecticide Imidacloprid of Zebrafish (Danio rerio) Gonads: A Histopathological Approach

Annales de Limnologie - International Journal of Limnology ◽

10.1051/limn/2021021 ◽

2021 ◽

Vol 57 ◽

pp. 23

Author(s):

Cansu Akbulut

Keyword(s):

Interstitial Fibrosis ◽

Periodic Acid ◽

Testicular Tissue ◽

Seminiferous Tubules ◽

Control Group ◽

Periodic Acid Schiff ◽

Histological Techniques ◽

Zona Radiata ◽

Structural Deterioration ◽

Neonicotinoid Insecticide

Neonicotinoids are the new class of insecticides that are high target specificity to insects. Imidacloprid is a neonicotinoid that is the most widely used insecticide in the world. As a result of its widespread use in agriculture, imidacloprid interferes with the aquatic system and threatens the aquatic environment. In this study, an investigation of the histopathological effects of imidacloprid on zebrafish gonads was aimed. Zebrafish were exposed to 9.5 mg/L, 19 mg/L, and 38 mg/L of imidacloprid for 5 days, considering the 96 h LC50 value. After dissecting the gonadal tissues, routine histological techniques were applied, and the tissues were stained with Periodic Acid-Schiff (PAS), Toluidine Blue (TB), and Haematoxylin and Eosin (H&E). Sections were examined under a light microscope. While normal gonad histology was observed in the control group, histopathological alternations such as degeneration and union in the seminiferous tubules, hypertrophy in spermatogenic and Leydig cells, and interstitial fibrosis were detected in testicular tissue of the experimental groups. In the ovarian tissues of the experimental groups, structural deterioration in oocytes, autolysis, increase in the number of atretic oocytes, vacuolization in cortical alveoli, thickening and curling in the zona radiata, and opening in the perifollicular layers were detected.

Download Full-text

Three-dimensional analysis of seminiferous tubules and spermatogenic waves in mice

Reproduction ◽

10.1530/rep-17-0391 ◽

2017 ◽

Vol 154 (5) ◽

pp. 569-579 ◽

Cited By ~ 8

Author(s):

Hiroki Nakata ◽

Takahiro Sonomura ◽

Shoichi Iseki

Keyword(s):

High Performance ◽

Average Length ◽

Periodic Acid ◽

3D Structure ◽

Three Dimensional ◽

Rete Testis ◽

Seminiferous Tubules ◽

Periodic Acid Schiff ◽

Adult Mice ◽

Branching Points

The aim of the present study was to reconstruct seminiferous tubules and analyze spermatogenic waves in seminiferous epithelia in developing and adult mice using serial paraffin sections and high-performance three-dimensional (3D) reconstruction software. By labeling the basement membrane of seminiferous tubules with fluorescent immunohistochemistry or periodic acid-Schiff-hematoxylin staining, all seminiferous tubules were reconstructed in 9 testes from 9 different mice, 3 each at 0, 21 and 90 days (adult) postpartum. The 3D structure of seminiferous tubules, including the number and length of tubules as well as the number of connections with the rete testis, branching points and blind ends, was assessed accurately. Although tubules showed marked variations among individual mice, their overall structure was regular and retained from newborn to adult mice. Some seminiferous tubules contained inner portions running distant from the testis surface. In a representative testis at 21 days, the sites at which spermatids initially occurred were examined by labeling acrosomes and were found to be preferentially distributed in the upper and medial portions of the testis close to the rete testis. In a representative adult testis, 76 complete waves with an average length of 16.9 mm were found and their directions were analyzed. The methods used in the present study will be useful for investigating the structure and function of seminiferous tubules in mice and humans under normal and pathological conditions, such as infertility.

Download Full-text

Granulomatous Epididymitis Related to Rhodotorula glutinis Infection in a Dog

Veterinary Pathology ◽

10.1177/030098589503200615 ◽

1995 ◽

Vol 32 (6) ◽

pp. 716-718 ◽

Cited By ~ 10

Author(s):

K. Kadota ◽

K. Uchida ◽

T. Nagatomo ◽

Y. Goto ◽

T. Shinjo ◽

...

Keyword(s):

Connective Tissue ◽

Sertoli Cells ◽

Periodic Acid ◽

Rhodotorula Glutinis ◽

Seminiferous Tubules ◽

Spermatogenic Cells ◽

Cell Debris ◽

Periodic Acid Schiff ◽

Microbiological Characteristics ◽

Cut Surface

A 4-year-old, male Great Dane dog developed severe swelling of the scrotum on 9 December 1991, and the testes and epididymides were removed surgically on 12 December 1992. The cut surface of the epididymides consisted of hard connective tissue and several small abcesses with slight hemorrhage. Histopathologically, the seminiferous tubules in the testes had only a few spermatogenic cells, but Sertoli cells were well preserved. Both epididymides consisted entirely of a proliferation of fibrous connective tissue, and only a few ducts deferens containing cell debris, neutrophils, and macrophages in the lumina were present. In all lesions of the epididymides, the macrophages contained periodic acid–Schiff– and Grocott's silver–positive round granules, 5-8 μm in diameter. Microbiologically, smooth salmon-pink colonies consisting of ovoidal yeast, about 10 μm in diameter, were isolated from the samples of epididymides but not from those of the testes. The isolated yeast had microbiological characteristics of Rhodotorula glutinis. From these observations, we diagnosed the present case as granulomatous epididymitis due to Rhodotorula infection.

Download Full-text

Gonadal development in males of the orange mud crab, Scylla olivacea (Herbst, 1796) (Decapoda, Brachyura, Portunidae)

Crustaceana ◽

10.1163/15685403-00003622 ◽

2017 ◽

Vol 90 (1) ◽

pp. 1-19 ◽

Cited By ~ 19

Author(s):

K. Waiho ◽

H. Fazhan ◽

S. Jasmani ◽

M. Ikhwanuddin

Keyword(s):

Germ Cells ◽

Reproductive System ◽

Vas Deferens ◽

Gonadosomatic Index ◽

Gonadal Development ◽

Gonad Maturation ◽

Mud Crab ◽

Maturation Stages ◽

Histological Characteristics

This study describes the morphological and histological characteristics of the male reproductive system of the orange mud crab, Scylla olivacea (Herbst, 1796). Three maturation stages were determined on the basis of the vas deferens diameter, the gonad colour and the gonadosomatic index (GSI). Significant differences were observed in the GSI value as the crabs matured. All germ cells were present in the histological sections of the testes, and these decrease significantly in size as they progress from spermatogonia, spermatocytes, and spermatids to spermatozoa. Spermatophores are found in the anterior vas deferens (AVD) and median vas deferens (MVD) of all maturation stages but not in the posterior vas deferens (PVD), with a significantly smaller size in immature and maturing specimens. Thus, the classification of S. olivacea gonads into various maturation stages based solely on histological characteristics (i.e., the presence of spermatophores) is impossible. Therefore, the combination of both microscopic and macroscopic features is essential for determining the gonad maturation stages of male S. olivacea.

Download Full-text

Identification of glycoproteins associated with elastin-associated microfibrils.

Journal of Histochemistry & Cytochemistry ◽

10.1177/33.4.3980982 ◽

1985 ◽

Vol 33 (4) ◽

pp. 287-294 ◽

Cited By ~ 18

Author(s):

J C Fanning ◽

E G Cleary

Keyword(s):

Lectin Binding ◽

Periodic Acid ◽

Periodate Oxidation ◽

Ruthenium Red ◽

Elastic Tissue ◽

Electron Microscopic ◽

Periodic Acid Schiff ◽

Mature Adult ◽

Aldehyde Groups ◽

Nuchal Ligament

The microfibrils associated with elastic tissue have been shown to be predominantly proteinaceous. On the basis of their affinity for cationic stains, including ruthenium red, they have been assumed to be glycoprotein, but more evidence to support this claim has not been adduced. Despite repeated investigation of glycoprotein materials obtained by extraction of elastic tissues with reagents that appear to remove microfibrils, the chemical composition of elastin-associated microfibrils remains obscure. An electron microscopic study of the microfibrils in two elastin-rich tissues (bovine nuchal ligament and aorta) during their development was pursued using more specific histochemical methods. The periodic acid-alkaline bismuth stain (analogous to the periodic acid-Schiff stain for glycoproteins in light microscopy) has been adapted for this study. Specific aldehyde groups (confirmed by blocking with m-aminophenol or sodium borohydride) were identified after periodate oxidation as fine granules of bismuth stain. These were shown to localize specifically along the elastin-associated microfibrils in a finely punctate form. Staining of the amorphous elastic component did not occur except for a fine rim adjacent to the microfibrils. Lectin binding with concanavalin A (with ferritin markers) confirmed that there are glucose- or mannose-containing proteins associated with the microfibrillar component of elastic tissue. This was true of these microfibrils in all layers of the aortic wall and throughout the ligament. It was also true of mature adult tissues in which there was a lesser proportion of microfibrils. It is concluded that elastin-associated microfibrils really are associated with glycoprotein(s).

Download Full-text

Transcriptomic and microstructural analyses in Liriodendron tulipifera Linn. reveal candidate genes involved in nectary development and nectar secretion

10.21203/rs.2.9840/v3 ◽

2019 ◽

Author(s):

Huanhuan Liu ◽

Jikai Ma ◽

Huogen Li

Keyword(s):

Candidate Genes ◽

Anthocyanin Biosynthesis ◽

Expression Profiles ◽

Periodic Acid ◽

Middle Part ◽

Pcr Analysis ◽

Rna Seq ◽

Nectar Secretion ◽

Relict Species ◽

Periodic Acid Schiff

Abstract Background: Nectar is a major floral attractant and reward for insects that ensures pollination. Liriodendron, a genus of the Magnoliaceae family, includes only two relict species, L. chinense and L. tulipifera, which are considered “basal angiosperms” according to plant evolutionary history. The flowers of Liriodendron plants are insect pollinated and secrete nectar to attract pollinators. To date, the morphology and anatomy of nectaries, the mechanism of nectar secretion and the molecular mechanism of nectary development in Liriodendron remain poorly understood. Methods: In this study, we examined the nectary surface cells and change in starch in L. tulipifera by using scanning electron microscopy and periodic acid-Schiff techniques to select appropriate samples for subsequent research. Transcriptome sequencing was of the top and middle parts of immature nectaries and the middle part of mature and postsecretory nectaries in L. tulipifera was performed. We evaluated the expression profiles of 21 DEGs that are closely related to nectary development and nectar secretion for real-time quantitative PCR analysis. Results: L. tulipifera nectaries are starch-storing nectaries and are located in the top and middle parts of L. tulipifera petals. After analyzing the RNA-seq data, we obtained 115.26 Gb of clean data in 12 libraries and mapped the results to the L. chinense reference genome with 71.02%-79.77% efficiency. In total, 26,955 DEGs were identified by performing six pairwise comparisons. The flavonoid biosynthesis, phenylpropanoid biosynthesis, anthocyanin biosynthesis and starch and sucrose metabolism pathways were enriched and related to nectar secretion and pigment change. We identified 56 transcription factor families, and members of the TCP, Trihelix, C2H2, ERF, and MADS families changed dynamically during nectary development. Moreover, to further verify the accuracy of the RNA-seq results, we validated the expression profiles of 21 candidate genes. Conclusions: We evaluated the nectary development and secretion processes comprehensively and identified many related candidate genes in L. tulipifera. These findings suggest that nectaries play important roles in flavonoid synthesis and petal color presentation.

Download Full-text

Characterization of a protandrous hermaphroditic reproductive system in transitional and female phases of Norileca indica — morphological, histological and ultrastructural approach

Crustaceana ◽

10.1163/15685403-00003896 ◽

2019 ◽

Vol 92 (6) ◽

pp. 641-664 ◽

Cited By ~ 1

Author(s):

Helna Ameri Kottarathil ◽

Sudha Kappalli

Keyword(s):

Reproductive System ◽

Vas Deferens ◽

Gonadosomatic Index ◽

Oocyte Size ◽

Reproductive Life ◽

Maturation Stages ◽

Female Phase ◽

Transitional Phase

Abstract The present study explores the reproductive system of Norileca indica during its transitional and female phase at morphological, histological, ultrastructural and histochemical levels. The paired and symmetrical hermaphroditic reproductive system of N. indica in the transitional and female phases lies dorsally in the thorax on either side of the gut, each consisting of a three-lobed testis (with lobes t1, t2 and t3) followed by an ovary and then a vas deferens, which opens into the paired penes located at sternite 7; the oviduct, arising laterally from the ovarian lobe, opens into the gonopore located on the 6th pereonite. In the transitional phase, the gonads show a presence of germ cells at different maturation stages: spermiogenesis in the testes has already halted, while the ovary undergoes active vitellogenesis. Spermatophores are frequently seen in the vas deferens but seldom in the testes; the size of the oocytes then is 250-1200 μm. The gonadosomatic index (GSI) and nucleocytoplasmic index (NCI) range over 0.090-0.198 and 0.46-0.11, respectively. In the female, oocyte size increases to 1500 μm; the GSI ranges 0.019-0.235 and the NCI from 2.40 to 0.09; testes and vas deferens are not prominent. This paper discusses the possible role of protandrous hermaphroditism in the reproductive life of N. indica.

Download Full-text

Antioxidant and cytoprotective effects of Nigella sativa L. seeds on the testis of monosodium glutamate challenged rats

Scientific Reports ◽

10.1038/s41598-021-92977-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mahmoud Abd-Elkareem ◽

Mokhless A. M. Abd El-Rahman ◽

Nasser S. Abou Khalil ◽

Ayman S. Amer

Keyword(s):

Nigella Sativa ◽

Periodic Acid ◽

Monosodium Glutamate ◽

Food Additives ◽

Lipid Peroxides ◽

Basement Membranes ◽

Seminiferous Tubules ◽

Control Group ◽

Albino Rats ◽

Periodic Acid Schiff

AbstractMonosodium glutamate (MSG) is one of the most widely spread food additives that might cause male infertility. However, Nigellasativa L. seeds (NSS) could provide a solution. This study was designed to investigate the potential effects of NSS on rats ingesting MSG. To achieve this aim, adult male albino rats were randomly equally assigned into three groups for 21 days: control group received no treatment, MSG group received MSG as 30 g/kg feed, and MSG + NSS group received MSG as 30 g/kg and NSS as 30 g/kg feed. Testis histomorphometry showed marked deterioration by MSG as atrophic seminiferous tubules with degeneration of their lining cells, damaged Leydig cells and decreased germ cells number. Periodic Acid Schiff stain indicated irregular interrupted basement membranes. Glutathione reductase, superoxide dismutase 2 (SOD2), and caspase-3 immuno-expressions increased in testicular cells. Testosterone levels were significantly decreased in MSG challenged rats along with significant increase in luteinizing hormone levels, whereas NSS normalized this hormonal profile. MSG exposure also caused significantly increased lipid peroxides (LPO), glutathione-S-transferase, and total antioxidant capacity (TAC) whereas nitric oxide and SOD2 were significantly decreased. NSS succeeded in rebalance LPO and TAC and ameliorated the histoarchitectural disturbances. NSS mitigated MSG-induced testicular impairment by its antioxidant and cytoprotective activities.

Download Full-text