We want to examine the effects of one chemotherapy medicine on mouse spermatogenesis and we need to compare the number of spermatids, spermatocytes, spermatogonia, Sertoli and Leydig cells between two groups. Can we use the paraffin sections for this purpose or we have to provide resin sections?
Thank you
Lab Anim Res. 2015 Sep; 31(3): 111–116.
Published online 2015 Sep 30. doi: 10.5625/lar.2015.31.3.111
PMCID: PMC4602077
Effects of pentoxifylline administration on histomorphological parameters of streptozotocin-induced diabetic rat testes
Abbas Piryaei,1 Azam Najar,1 and Mohammad Bayat 2
Author information ► Article notes ► Copyright and License information ►
Go to:
Abstract
The effect of pentoxifylline (PTX) administration on histomorphological parameters of streptozotocin (STZ)-induced type 1 diabetes mellitus (DM) in male rat testes were evaluated. We randomly divided 40 male rats into the following four groups: group 1: control or normal glycemic (NG) rats; group 2 or NG rats that received only normal saline (NS), (NG+NS); group 3 or diabetic rats which were not treated by PTX (DM+vehicle solution (NS)); and group 4 which comprised diabetic rats treated with 50 mg/kg of PTX (DM+PTX). Type 1 DM was induced by intraperitoneal injection of STZ (55 mg/kg). Rats were held for 30 days after which the experimental group received PTX twice daily (25 mg/kg) or NS. After 14 days of treatment by PTX or NS, the left testes from all rats were extracted and prpared for histological study. Apoptotic cells, blood vessel density, and spermatogenesis were evaluated. Data were analyzed by ANOVA test. PTX-treated-diabetic rats showed a significant decrease in number of apoptotic cells and decrease in blood vessel density compared to the DM+NS rats. A significant increase in spermatogenesis was observed in the PTX-treated diabetic group, compared to the DM+NS groups. It was concluded that PTX administration to STZ-induced type 1 DM rats affected apoptotic cell number positively. Moreover, blood vessel density significantly decreased and improvements were observed in spermatogenesis.
Keywords: Diabetes mellitus, pentoxifylline, histopathology, testes, rat
I suggest using sessions in resin with a thickness of the histological section of 2 µm. The resin enables to make cuts with smaller thicknesses than paraffin, as well as better staining as periodic acid – Schiff (PAS) + Iron Hematoxylin + Metanil Yellow (QUINTERO-HUNTER; GRIER; MOSCATO, Biotech Histochem. 1991; 66(4):169-72), for example, than Hematoxylin and Eosin, showing better the details of cell structures of the testes. The thickness of 2 µm of the resin's histological sections improves sharpness by reducing the appearance of overlapping structures. I consider appropriate using this technique as a tool for the morphometric and stereological study of testes, however, you must first standardize the thickness of histological sections measuring larger structures to be studied in order to avoid counting twice the same structure.
in my opinión it does not matter that much whether you use paraffin or platic sections as son as they are GOOD SECTIONS of know thincknes. The thickness to use will depend on what are you trying to quantify.
We have used stereology for many studies. I am copying the references for some papers we publish using these techiques. In the old ones we simply counted cells and corrected the counts with available formulas. In more cent ones we used image analysing systems that give a lot more of possiblities when expressing your results. It all depends on what do you have and how hard you are willing to work.
good luck
these are the references:
R A Rey, S M Campo, P Bedecarrás, C Nagle, H E Chemes. Is infancy a quiescent period of testicular development? Histologic, morphometric and functional study of the seminiferous tubules of the cebus monkey from birth to the end of puberty. J Clin Endocr Metab 76:1325-1331, 1993.
R A Rey, C Nagle, H E Chemes. Morphometric study of the testicular intersticial tissue of the monkey cebus apella during postnatal development. Tissue and Cell 28(1), 31-42, 1996.
Chemes HE, Rey RA, Nistal M, Regadera J, Musse M, Gonzalez Peramato P, Serrano A. Physiological Androgen Insensitivity of the Fetal, Neonatal and Early Infantile Testis is Explained by the Ontogeny of the Androgen Receptor Expression in Sertoli Cells. J. Clin. Endocrinol. Metab. 93: 4408-4412, 2008.
Quintana S, Venara M, Rey R, Clemente N, Chemes HE. Origin and evolution of somatic cell testicular tumours in transgenic mice. J. Pathol. 2010: 221 (4), 443-451.
- Badges
- Science topic
- Similar topics
- Philosophy
- Philosophy Of Science
- Reasoning