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雄性大鼠的生殖母细胞在出生后恢复迁移活性。

Gonocytes of male rats resume migratory activity postnatally.

作者信息

McGuinness M P, Orth J M

机构信息

Department of Anatomy and Cell Biology, Temple University School of Medicine, Philadelphia, PA.

出版信息

Eur J Cell Biol. 1992 Oct;59(1):196-210.

PMID:1361432
Abstract

In the testis of the neonatal rat, maturation of germ cells, or gonocytes, lays the foundations for spermatogenesis which will begin later in postnatal development. One of the most critical and yet least understood of the events that occur during the immediate neonatal period is relocation of gonocytes from the more central part of the seminiferous cord, where they are surrounded by Sertoli cells, to its periphery, where they contact the basement membrane. For the current study, we examined this change in gonocyte position by identifying some of the cellular mechanism involved, with the aim of determining whether movement of gonocytes to the basement membrane in vivo and development of cellular processes by these cells in vitro represents a resumption of migratory activity similar to that displayed by their fetal ancestors and by other motile cells. First, we used either thiamine pyrophosphatase cytochemistry or the fluorescent probe nitrobenzoxadiazole ceramide to visualize the Golgi complex in gonocytes and found that (1) this organelle matures and apparently enlarges in vivo with a time course paralleling movement of gonocytes to the basement membrane and undergoes similar changes in vitro that correlate with gonocyte process formation, and (2) the Golgi complex is located in perinuclear cytoplasm facing the apparent direction of gonocyte movement in vivo and in cytoplasm near the cellular process in the great majority of elongated gonocytes in coculture. Next we used two drugs, brefeldin A and monensin, which have in common their ability to disrupt the Golgi complex, and found that both drugs prevent process formation by gonocytes in a manner that is completely reversible. We also tested the involvement of the cytoskeleton in gonocyte elongation by utilizing nocodazole to disrupt and taxol to stabilize microtubules, as verified by alpha-tubulin immunofluorescence. Inclusion of the drug abolished (taxol) or substantially diminished (nocodazole) the ability of gonocytes to elongate in a reversible manner. We also found that the Golgi complex was intact in the presence of taxol and that microtubules were intact in the presence of both Golgi complex-specific drugs. Thus, our findings indicate that (1) both the Golgi complex and microtubules are involved in development of processes by gonocytes and (2) neither structure is sufficient by itself to allow these cells to elongate. Taken together, our data provide new evidence suggesting that the cellular mechanism utilized by postnatal gonocytes in relocating to the basement membrane are those mediating active migration.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

在新生大鼠的睾丸中,生殖细胞即生殖母细胞的成熟为稍后在出生后发育阶段开始的精子发生奠定了基础。在新生儿早期发生的诸多事件中,最关键却又最不为人所理解的事件之一,是生殖母细胞从生精索较中央的部分(在那里它们被支持细胞所包围)迁移到其外周,在那里它们接触基底膜。在本研究中,我们通过确定一些相关的细胞机制来研究生殖母细胞位置的这种变化,目的是确定生殖母细胞在体内向基底膜的移动以及这些细胞在体外形成细胞突起是否代表着类似于其胎儿祖先和其他运动细胞所表现出的迁移活性的恢复。首先,我们使用硫胺素焦磷酸酶细胞化学或荧光探针硝基苯并恶二唑神经酰胺来观察生殖母细胞中的高尔基体复合体,发现:(1)该细胞器在体内成熟并明显增大,其时间进程与生殖母细胞向基底膜的移动平行,并且在体外经历类似的变化,这些变化与生殖母细胞突起的形成相关;(2)在体内,高尔基体复合体位于核周细胞质中,朝向生殖母细胞移动的明显方向,在共培养的绝大多数伸长的生殖母细胞中,高尔基体复合体位于靠近细胞突起的细胞质中。接下来,我们使用了两种药物,布雷菲德菌素A和莫能菌素,它们的共同特点是能够破坏高尔基体复合体,并且发现这两种药物都以完全可逆的方式阻止生殖母细胞形成突起。我们还通过利用诺考达唑破坏微管和紫杉醇稳定微管来测试细胞骨架在生殖母细胞伸长中的作用,α-微管蛋白免疫荧光证实了这一点。加入该药物消除了(紫杉醇)或显著降低了(诺考达唑)生殖母细胞以可逆方式伸长的能力。我们还发现,在存在紫杉醇的情况下高尔基体复合体是完整的,在存在两种高尔基体复合体特异性药物的情况下微管是完整的。因此,我们的研究结果表明:(1)高尔基体复合体和微管都参与了生殖母细胞突起的形成;(2)这两种结构单独都不足以使这些细胞伸长。综上所述,我们的数据提供了新的证据,表明出生后生殖母细胞迁移到基底膜所利用的细胞机制是介导主动迁移的机制。(摘要截短至400字)

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