Visualization of Reinke’s crystals in normal and cryptorchid testis

Abstract

Within the human testis, Reinke's crystals are found in Leydig cells but their nature and function are poorly understood. The aim of our study was to investigate the properties of Reinke's crystals in man with the normal morphology of the testis (control group) and infertile patients diagnosed with cryptorchidism. 20 biopsies from infertile patients and six biopsies from men with regular spermatogenesis (20-30 years.) were used. Sections of the testis tissue were stained with haematoxylin and eosin and a modified Masson's method. Specimens were observed by bright field, confocal and transmission electron microscopy (TEM). The number of Reinke's crystals in investigated groups was determined applying stereological methods. In both groups, Reinke's crystals were noted within the cytoplasm and nuclei of Leydig cells. Some "free" crystals were found within the interstitial space, outside Leydig cells. Confocal microscopy proved to be very useful in the assessment of the shape and 3D reconstruction of the crystal. TEM analysis confirmed a hexagonal form of the crystal, while crystallographic data on sections of 70-300 nm thickness provided a better insight into the organization of the crystal lattice. Stereological analysis revealed a significant increase in the number of crystals in cryptorchid testes when compared with controls. Increased number of crystals in cryptorchid specimens leads to the assumption that the prolonged exposure to higher (abdominal) temperature might stimulate enzymes involved in the synthesis of the proteins of the crystal. However, the exact molecular nature of the crystal lattice remains in both normal and cryptorchid testis obscure.

Leydig cell clustering and Reinke crystal distribution in relation to hormonal function in adult patients with testicular dysgenesis syndrome (TDS) including cryptorchidism

“…[28][29][30][31] In this study we re-investigated a previously reported finding that the number of RCs in undescended testes may be significantly greater than in normally descended testes. 15 We have confirmed a high number of RC-containing LCs in patients with a history of undescended testes, and revealed an interesting pattern of an increasing proportion of the RC-containing cells with the more cranial testis position and with significantly greater abundance of RCs in intra-abdominal testes. This is in line with influence of temperature, suggested by Kozina's study.…”

“…This is in line with influence of temperature, suggested by Kozina's study. 15 Surprisingly, in our study testes that were cryptorchid at birth, and especially unilaterally undescended testes, seemed to contain higher numbers of RCs as compared to normally descended testes in TDS patients. It was unexpected because the specimens from men with a history of cryptorchidism often contain LC micronodules.…”

“…[11][12][13] In crystallographic and electron microscopy studies, the crystals are visualized as hexagonal or polygonal prisms with an interior honeycomb pattern composed of 50 Å microfilaments, consistent with a proteinaceous content. [14][15][16] A recent study provided evidence that the crystals stain specifically with antibodies for 3β-hydroxysteroid dehydrogenase (β-HSD), which is a key enzyme needed for biosynthesis of all steroid hormones. 16 RCs have been reported to be absent in case studies of patients with Down's syndrome, androgen insensitivity syndrome and 17β-HSD deficiency.…”

Reinke´s Crystals in Perivascular and Peritubular Leydig Cells

“…Their arrangement seems to support building of a larger crystal from smaller, immature forms. 39 In specimens originating from control biopsies we were able to establish a certain difference in number of Reinke´s crystals in perivascular and peritubular Leydig cells. Although not statistically significant, our stereological data indicated a lower number of crystals in peritubular cells.…”

“…In order to calculate the above-mentioned relative variable, appropriate formulae for numerical density have been applied. [35][36][37][38][39] The total number of crystals was determined by multiplying the testis volume (determined by the ultrasound examination) and numerical density. …”