Heterogeneous nuclear ribonucleoprotein L (hnRNP L) is a major pre-mRNA binding protein and it is an abundant nuclear protein that shuttles between the nucleus and the cytoplasm. hnRNP L is known to be related to many cellular processes, including chromatin modification, pre-mRNA splicing, mRNA export of intronless genes, internal ribosomal entry site (IRES)-mediated translation, mRNA stability, and spermatogenesis. In order to identify the cellular proteins interacting with hnRNP L, this study performed a yeast two-hybrid screening, using a human liver cDNA library. The study identified insulin-like growth factor binding protein-4 (IGFBP-4) as a novel interaction partner of hnRNP L in the human liver. It then discovered, for the first time, that hnRNP L interacts specifically with IGFBP-4 in a yeast two-hybrid system. The authenticity of this two-hybrid interaction of hnRNP L and IGFBP-4 was confirmed by an in vitro pull-down assay.
In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the Sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ${\mu}$m in length. The round head was 2.2 ${\mu}$m long. The middle piece developed beneath the head was 0.5 ${\mu}$m long. Tail was 40 ${\mu}$m in length. Coomassie brilliant blue (CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.
Park, Young-Je;Chung, Ee-Yung;Lee, Jeong-Yong;Park, Kwang-Jae
The Korean Journal of Malacology
/
v.22
no.1
s.35
/
pp.39-49
/
2006
Ultrastructure of germ cell differentiation during supermatogenesis and the reproductive cycle in male Patinopecten yessoensis was studied by histological and cytological observations. The gonadosomatic index (GSI) in males rapidly increased and reached a maximum in April when seawater temperature gradually increased. Then the GSI gradually decreased from May through July when spawning occurred. Accordingly, monthly changes in the GSI in males coincided with testicular maturation and spawning periods. The sperm morphology of P. yessoensis belongs to the primitive type and showed general characteristics of external fertilization species. The head of the spermatozoon is approximately $3.50{\mu}m$ in length: the sperm nucleus and acrosome are approximately $2.90{\mu}m\;and\;0.60{\mu}m$ in length, respectively. The nuclear type of the spermatozoon is vase in shape, and the acrosome is cone type. The axoneme of the tail flagellum consists of nine pairs of microtubules at the periphery and a pair of central microtubules in the center The satellite body (which is formed by the centriole) and four mitochondria appear in the middle piece of the spermatozoon. The spawning period was from April through July and the main spawning occurred from May to June when seawater temperatures gradually increased. The reproductive cycle of this species can be classified into five successive stages; early active stage (September to November), late active stage (October to March), ripe stage (February to August), spawning stage (April to July), and spent/inactive stage (July to November).
Panax ginseng (family- Araliaceae) is a native plant of Korea and has been used for past several years among oriental people. To evaluate the radioprotective potential of P. ginseng on the formation of giant cells in the testis of Swiss albino mice, the animals were divided into four groups: -(I)-Only vehicle was administered. (II)P. ginseng treated group: -The animals received 10 mg/kg body weight P. ginseng root extract (in DDW) i.p. continuously for 30 days. (III) Irradiated group: -The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. (IV) Combined treatment group: -Animals were given P. ginseng extract for four days and on fourth day they were irradiated to 8 Gy gamma radiation after 30 minute of extract administration. The animals of these three groups were autopsied on day 1,3, 7, 14 and 30 days. In ginseng treated group, active spermatogenesis was observed without any toxic effect. Histopathological studies of irradiated group (II) revealed reduction in germ cell count, loss of sperms and formation of multinucleated giant cells on day 7th. These giant cells were formed by round nuclei of early or late spermatids. In combination group (III), although germinal epithelium was still disorganized with loss of cells in few tubules, but no giant cell formation was observed. In order to know the mechanism of radioprotection of ginseng, LPO and GSH were estimated. It was observed that pretreated irradiated animals showed inhibition of LPO and increase in GSH. Thus the present study suggests ginseng protects male gonads. This may be attributed to the inhibition of LPO and increase synthesis of GSH byginseng.
The formation of the acrosome during spermatogenesis in Gerris paludum was studied. The Golgi bodies are dispersed randomly in the cytoplasm at the early stage of the spermatocyte and get together to form several group of many bodies, and then they are equally divided into the spermatids by the meiotic divisions. The acroblast first appears in the form of a vesicle and soon an acrosomal granule is differentiated within it. The acroblast is separated from the acrosomal granule at the posterior of the nucleus and is finally sloughed off along the tail filament. The acrosome, after moving to the side of the nucleus opposite the mitochondrial derivatives, differentiates into two zones. The two basal bodies and the differentiated tip originate from the sheath. The basal bodies appear at the proximal part of the sheath simply in contact with the core on one side. During elongation and and narrowing of the acrosomes of the spermatids, they surround the one side at the base of the acrosome and finally all the other are immediately adjacent to the nucleus. The differentiated tip continues to the sheath at the anterior of the cores and is elongated prior to the two basal bodies. They appear to be contiguous twin-tubes, not a single granule in the later stage of the spermatids, and a group of the basal bodies in the sperm bundle.
Closely correlated expression patterns between ubiquitin specific peptidase 9X-linked (USP9X) and adherens junction formation factor (Afadin) in mouse testis development suggests that Usp9x regulates the deubiquitination of Af-6 (also known as Afadin, AFDN), and subsequently, the cell adhesion dynamics during gametogenesis. However, this relationship has not yet been tested in other domestic animals. The study was examined the temporal and spatial expression patterns of porcine USP9X and AFDN from the pre-pubertal to adult stages using real time-PCR and immunohistochemistry. Furthermore, we detected the transcripts of USP9X and AFDN in the testis of 1-, 6- and 12-months old boar, respectively. USP9X and AFDN were found to have similar expressions patterns, with basal expression after 1 month followed by a significant up-regulation from 6 months (puberty) onwards. In addition, neither the AFDN or USP9X proteins were detected in spermatogenic cells but they were expressed in the leydig cells and sertoli cells. USP9X was detected around the basal lamina during pre-puberty, and predominantly expressed in the leydig cells at puberty. Finally, in adult testis, USP9X was increased at the sertoli cell-cell interface and the sertoli cell-spermatid interface. In summary, closely correlated expression patterns between USP9X and AFDN in boar testis supports the previous findings in mice. Furthermore, the junction connections between the sertoli cells may be regulated by the ubiquitination process mediated via USP9X.
Acetaminophen [Paracetamol, N-acetyl-para-aminophenol (APAP)] is a common over-the-counter analgesic agent as nonsteroidal anti-inflammatory drugs (NSAIDs). The high doses or the long-term treatment of acetaminophen via usual gavage feeding resulted in damage of testicles that presented recoverable impairment, as well as liver and kidney. The influence of acetaminophen was examined in male golden hamsters treated with acetaminophen-containing diet feeding. They were divided into 5 groups and subjected to this experiment for 4 weeks: animals housed in long photoperiod (LP) as LP control, animals housed in short photoperiod (SP) for 4 weeks as SP control (SP4), and groups of animals treated with low, middle, and high concentrations of acetaminophen (Low, Middle, High groups). Also animals housed in SP for 8 weeks were included (SP8) to contrast testicular activities, if necessary. As results, spermatozoa filled the seminiferous tubules of the testicles of animals in LP control and SP4 groups. The aspects were seen in the animals taken diets of low and middle doses of acetaminophen. The animals who fed high dose of acetaminophen showed large or small testicles. The large testicles displayed all germ cells at the steps of spermatogenesis. The small testicles presented no sperm as the animals housed in SP for 8 weeks. Thus these results indicate that acetaminophen invokes the antigonadal effects and accelerates the regressing process of the testicles in the animals compared to the animals exposed to SP.
Objective: Testicular fat deposition has been reported to affect animal reproduction. However, the underlying mechanism remains poorly understood. The present study explored whether sperm meiosis and testosterone synthesis contribute to mouse testicular fat deposition-induced reproductive performance. Methods: High fat diet (HFD)-induced obesity CD1 mice (DIO) were used as a testicular fat deposition model. The serum hormone test was performed by agent kit. The quality of sperm was assessed using a Sperm Class Analyzer. Testicular tissue morphology was analyzed by histochemical methods. The expression of spermatocyte marker molecules was monitored by an immuno-fluorescence microscope during meiosis. Analysis of the synthesis of testosterone was performed by real-time polymerase chain reaction and reagent kit. Results: It was found that there was a significant increase in body weight among DIO mice, however, the food intake showed no difference compared to control mice fed a normal diet (CTR). The number of offspring in DIO mice decreased, but there was no significant difference from the CTR group. The levels of follicle-stimulating hormone were lower in DIO mice and their luteinizing hormone levels were similar. The results showed a remarkable decrease in sperm density and motility among DIO mice. We also found that fat accumulation affected the meiosis process, mainly reflected in the cross-exchange of homologous chromosomes. In addition, overweight increased fat deposition in the testis and reduced the expression of testosterone synthesis-related enzymes, thereby affecting the synthesis and secretion of testosterone by testicular Leydig cells. Conclusion: Fat accumulation in the testes causes testicular cell dysfunction, which affects testosterone hormone synthesis and ultimately affects sperm formation.
The epididymal fat is a type of gonadal adipose tissue, which is localized closely to the testis. Even though it has been suggested that the epididymal fat is necessary for maintenance of spermatogenesis in the testis, the influence of epididymal fat on expression of testicular steroidogenic enzymes has not been examined. In the present research, expressional changes of steroidogenic enzymes in the mouse testis after 2 weeks of the surgical partial lipectomy of epididymal fat at different postnatal ages were determined by real-time polymerase chain reaction analysis. The transcript levels of all molecules at 2 months of postnatal age were significantly increased by the lipectomy of epididymal fat. However, the lipectomy at 5 months of postnatal age resulted in decreases of expression levels of all molecules examined in the testis. Except a reduced transcript level of hydroxysteroid 17-beta dehydrogenase 3, there were no significant changes of expression levels of other steroidogenic enzymes by the lipectomy at 8 months of postnatal age. At 12 months of postnatal age, the lipectomy caused a significant increase of transcript level of steroidogenic acute regulatory protein and a significant decrease of transcript level of hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1, without any expressional change of cytochrome P450 side chain cleavage, hydroxysteroid 17-beta dehydrogenase 3, and hydroxysteroid 17-beta dehydrogenase 3 in the testis. These findings suggest that the substances derived from epididymal fat could differentially influence on expression of steroidogenic enzymes in the testis during postnatal period.
Kim, Jong-choon;Lee, Hyun-sook;Yun, Hyo-in;Chung, Moon-koo
Korean Journal of Veterinary Research
/
v.40
no.2
/
pp.361-371
/
2000
It has been recently reported that 2-bromopropane (2-BP) induces male reproductive toxicity in both human and experimental animals. However, delayed effects of 2-BP on male reproductive system have not been investigated in detail. The present study was conducted to investigate the testicular toxicity of 2-BP and to determine the recovery of normal spermatogenesis in Sprague-Dawley rats. Male rats aged 5 weeks were administered 1,000mg/kg 2-BP by gavage daily for 4 weeks and sacrificed sequentially at 1, 2, 3, 4 and 12 weeks after initiation of 2-BP treatment. Testicular toxicity was evaluated qualitatively by histopathological examinations and quantitatively by reproductive organ weights, spermatid head count, and repopulation index. In the 2-BP treated rats, the body weights was significantly suppressed and the weights of testes and epididymides were also decreased in a time-dependent manner. On histopathological examination, spermatogonia in stages I-VI and preleptotene and leptotene spermatocytes in stages VII-IX were strongly depleted at 1 week of dosing. Spermatogonia were depleted extensively in all spermatogenic stages at 2 weeks. Continuing with the evolution of spermatogenic cycle, zygotene spermatocytes, pachytene spermatocytes, and round spermatids were sequentially depleted at 2, 3, and 4 weeks of dosing due to the depletion of their precursor cells. Vacuolization of Sertoli cells and spermatid retention were also observed at all time points, suggesting that 2-BP induced Sertoli cell dysfunction. At 12 weeks, after 8 weeks recovery, most of the tubules appeared severely atrophic and were lined by Sertoli cells only. Leydig cell hyperplasia in the interstitial tissue was also found. In addition, dramatic reductions in the number of spermatid heads and repopulation index were observed, indicating that 2-BP-induced testicular injury is irreversible. These results indicate that 4 weeks repeated-dose of 1,000mg/kg 2-BP results in a progressive germ cell loss due to the depletion of spermatogonia followed by long-term testicular atrophy in SD rats.
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