Spermatogenesis in the golden hamster during the first spermatogenic wave: A flow cytometric analysis

Golan, Rachel; Weissenberg, R.; Oschry, Yitzchak; Shochat, L.; Lewin, L.

doi:10.1002/(sici)1098-2795(200002)55:2<205::aid-mrd10>3.0.co;2-p

Cited by 9 publications

(9 citation statements)

References 27 publications

(29 reference statements)

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“…Previous studies have investigated the expression profile of PLCz mRNA, showing that in mice PLCz mRNA was detectable in spermatids and not in testicular cells depleted of spermatids (7), whereas a more systematic study of PLCz mRNA expression during spermatogenesis in mice and pigs concluded that PLCz mRNA was first present in the round spermatid stage and most likely translated in elongated spermatids (10). In the present study we carried out Northern analysis of testes from a time-course of postnatal hamsters to study PLCz mRNA expression during spermatogenesis and found that PLCz mRNA was present at day 17, which is when meiotic spermatocytes first appear in this species (37). This apparent earlier pattern of expression of PLCz mRNA compared with previous reports may reflect species differences, as well as the fact that we used Northern blotting rather than RT-PCR to analyze expression.…”

Section: Plcz Mrna Is First Expressed In Spermatocytes In Hamstersmentioning

confidence: 67%

“…3B). Northern analysis was also used to study expression of PLCz mRNA during spermatogenesis in the hamster, using testicular tissue obtained from animals at 0, 5, 10, 17, 30, and 63 days after birth, these time points being based on a previous study that determined the time frame for the appearance of specific types of germ cells in neonate hamsters (37). We found that hamster PLCz mRNA could not be detected in testicular tissue until day 17 onward (Fig.…”

Section: Tissue Specificity and Expression During Spermatogenesis Of mentioning

confidence: 84%

“…We found that hamster PLCz mRNA could not be detected in testicular tissue until day 17 onward (Fig. 3C), corresponding to the time that early meiotic spermatocytes first appear (37).…”

Section: Tissue Specificity and Expression During Spermatogenesis Of mentioning

confidence: 95%

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Phospholipase C zeta undergoes dynamic changes in its pattern of localization in sperm during capacitation and the acrosome reaction

Young

Grasa

Coward

et al. 2009

Fertility and Sterility

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Section: Plcz Mrna Is First Expressed In Spermatocytes In Hamstersmentioning

confidence: 67%

Section: Tissue Specificity and Expression During Spermatogenesis Of mentioning

confidence: 84%

See 1 more Smart Citation

Phospholipase C zeta undergoes dynamic changes in its pattern of localization in sperm during capacitation and the acrosome reaction

Young

Grasa

Coward

et al. 2009

Fertility and Sterility

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“…For this purpose, we selected the first spermatogenic wave of the rat as a reference and compared, assuming proportionality to rat, expected length (EL) vs. observed length (OL) for the other two species [i.e., mouse (40) and guinea pig]. Besides, available data of Mesocricetus auratus (golden hamster) was also included for this comparison (45) (Fig. 3).…”

Section: Characterization Of Testis Postnatal Development In C Porcementioning

confidence: 99%

High‐purity flow sorting of early meiocytes based on DNA analysis of guinea pig spermatogenic cells

et al. 2011

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Mammalian spermatogenesis is still nowadays poorly understood at the molecular level. Testis cellular heterogeneity is a major drawback for spermatogenic gene expression studies, especially when research is focused on stages that are usually very short and poorly represented at the cellular level such as initial meiotic prophase I (i.e., leptotene [L] and zygotene [Z]). Presumably, genes whose products are involved in critical meiotic events such as alignment, pairing and recombination of homologous chromosomes are expressed during the short stages of early meiotic prophase. Aiming to characterize mammalian early meiotic gene expression, we have found the guinea pig (Cavia porcellus) as an especially attractive model. A detailed analysis of its first spermatogenic wave by flow cytometry (FCM) and optical microscopy showed that guinea pig testes exhibit a higher representation of early meiotic stages compared to other studied rodents, partly because of their longer span, and also as a result of the increased number of cells entering meiosis. Moreover, we have found that adult guinea pig testes exhibit a peculiar 4C DNA content profile, with a bimodal peak for L/Z and P spermatocytes that is absent in other rodents. Besides, we show that this unusual 4C peak allows the separation by FCM of highly pure L/Z spermatocyte populations aside from pachytene ones, even from adult individuals. To our knowledge, this is the first report on an accurate and suitable method for highly pure early meiotic prophase cell isolation from adult mammals, and thus sets an interesting approach for gene expression studies aiming at a deeper understanding of the molecular groundwork underlying male gamete production. ' International Society for Advancement of CytometryKey terms meiosis; spermatogenesis; flow cytometry; cell sorting; guinea pig; meiotic prophase; gene expression SPERMATOGENESIS can be viewed as the simultaneous and coordinated execution of three individual programs of gene expression: somatic proliferation of spermatogonia, meiosis, and spermiogenesis (1). In the seminiferous tubules of adult mammals, germ cells in different maturation steps coexist, with C (round spermatids, elongating and elongated spermatids, spermatozoa), 2C (several types of G1 spermatogonia, secondary spermatocytes) and 4C (different stages of primary spermatocytes, G2 spermatogonia) DNA content. Moreover, somatic Sertoli cells (2C) coexist with germ cells inside the tubules, which are surrounded by peritubular myoid cells and immersed in a stroma containing fibroblasts, lymphocytes, mastocytes, macrophages, and Leydig cells (all of them 2C).Beside the obvious importance of spermatogenesis as a source of male reproductive cells, along the lengthy first meiotic prophase significant events such as the recognition, alignment, pairing (synapsis), and recombination (crossing over) of homologous chromosomes take place (2-5). Particularly, meiotic homologous recombination has vital importance for genetic variability (6). Furthermore, mutations of

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“…The experimental interventions in the different groups were as follows: 1) the testis was merely removed from the body cavity; 2) the testis was removed and injected with a vehicle solution; 3) the testis was removed and injected with a vehicle solution and electroporated; 4) the testis was removed and electroporated without injection. Animals were left for 40 days following the procedure, which should allow, under normal circumstances, for a complete renewal of the spermatogenic cells in the testis; the spermatogenic cycle in the hamster is estimated to take 35 days [24].…”

Section: Effect Of In Vivo Gene Transfer On Testicular Integrity and mentioning

confidence: 99%

In Vivo Gene Transfer by Electroporation Allows Expression of a Fluorescent Transgene in Hamster Testis and Epididymal Sperm and Has No Adverse Effects upon Testicular Integrity or Sperm Quality1

Hibbitt

Coward

Kubota

et al. 2006

Biology of Reproduction

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The study of gene function in testis and sperm has been greatly assisted by transgenic mouse models. Recently, an alternative way of expressing transgenes in mouse testis has been developed that uses electroporation to introduce transgenes into the male germ cells. This approach has been successfully used to transiently express reporter genes driven by constitutive and testis-specific promoters. It has been proposed as an alternative method for studying gene function in testis and sperm, and as a novel way to create transgenic animals. However, the low levels and transient nature of transgene expression that can be achieved using this technique have raised concerns about its practical usefulness. It has also not been demonstrated in mammals other than mice. In this study, we show for the first time that in vivo gene transfer using electroporation can be used to express a fluorescent transgene in the testis of a mammal other than mice, the Syrian golden hamster. Significantly, for the first time we demonstrate expression of a transgene in epididymal sperm using this approach. We show that expression of the transgene can be detected in sperm for as long as 60 days following gene transfer. Finally, we provide the first systematic demonstration that this technique does not lead to any significant long-term adverse effects on testicular integrity and sperm quality. This technique therefore offers a novel way to study gene function during fertilization in hamsters and may also have potential as a way of creating transgenic versions of this important model species.

show abstract

Spermatogenesis in the golden hamster during the first spermatogenic wave: A flow cytometric analysis

Cited by 9 publications

References 27 publications

Phospholipase C zeta undergoes dynamic changes in its pattern of localization in sperm during capacitation and the acrosome reaction

Phospholipase C zeta undergoes dynamic changes in its pattern of localization in sperm during capacitation and the acrosome reaction

High‐purity flow sorting of early meiocytes based on DNA analysis of guinea pig spermatogenic cells

In Vivo Gene Transfer by Electroporation Allows Expression of a Fluorescent Transgene in Hamster Testis and Epididymal Sperm and Has No Adverse Effects upon Testicular Integrity or Sperm Quality1

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