Stereological and morphological analysis of the effects of aging on spermatogenesis in rat testis

Miranda, Eduardo Pereira de; Lorenzini, Fernando; Neves, Bruno Vinicius Duarte; Melchioretto, Eduardo Felippe; Hota, Thiago; Fraga, Rogerio De

doi:10.1590/s0102-865020180100000005

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Acta Cirúrgica Brasileira

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ORIGINAL ARTICLES, 5-Experimental Surgery • Acta Cir. Bras. 33 (10) • Oct 2018 • https://doi.org/10.1590/s0102-865020180100000005 copy

Stereological and morphological analysis of the effects of aging on spermatogenesis in rat testis ¹ 1 Research performed at Laboratory of Anatomy and Experimental Surgery, Division of Urology, Department of Surgery, Universidade Federal do Paraná (UFPR), Curitiba-PR, Brazil. Part of Master degree thesis, Postgraduate Program in Surgical Clinic. Tutors: Rogério de Fraga and Fernando Lorenzini.

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Purpose:

To quantify, through stereological and morphometric analysis, spermatogenesis in rats undergoing the natural aging process.

Methods:

Seventy-two male Wistar rats were divided into 6 equal groups according to age at the time of killing: 3, 6, 9, 12, 18, and 24 months. All the rats were subjected orchiectomy and collection of testicular parenchymal fragments for histological and morphometric analysis. The numerical density of spermatids was calculated using a stereological study, and morphometric analysis was conducted to measure the height of the germinal epithelium and the area of the seminiferous tubules.

Results:

We found that the 18 and 24 months groups showed a significant reduction in the number of round spermatids. However, the height of the germinal epithelium was not significantly different between the groups. The area of seminiferous tubules was also significantly reduced in the elderly rats compared to that in the young ones.

Conclusion:

Aging of rats showed a significant reduction in the number of round spermatids and the area of the seminiferous tubules, more pronounced in the rats at 18 and 24 months of life.

Key words:
Aging; Spermatogenesis; Infertility; Testis; Spermatids; Rats

Groups	N	Mean	SD	95% CI
3 months	360	75.74	15.96	74.08-77.39
6 months	360	71.75	17.77	69.91-73.59
9 months	360	72.13	17.63	70.30-73.95
12 months	360	72.60	17.37	70.79-74.39
18 months	360	28.64*	1.67	28.46-28.81
24 months	360	27.89*	6.17	27.24-28.52

Comparison		Difference between mean numerical densities of round spermatids	P value	95% CI
Group	Groups
3 months	6 months	3.98	0.003*	0.94-7.01
	9 months	3.60	0.009*	0.57-6.64
	12 months	3.13	0.038*	0.10-6.17
	18 months	47.09	0.000*	44.05-50.13
	24 months	47.88	0.000*	44.81-50.88
6 months	9 months	- 0.37	0.999	-3.40-2.66
	12 months	- 0.84	0.964	-3.87-2.19
	18 months	43.11	0.000*	40.07-46.15
	24 months	43.86	0.000*	40.83-46.90
9 months	12 months	-0.46	0.998	-3.50-2.56
	18 months	43.48	0.000*	40.45-46.52
	24 months	44.24	0.000*	41.20-47.27
12 months	18 months	43.95	0.000*	40.92-46.99
12 months	24 months	44.70	0.000*	41.67-47.74
18 months	24 months	0.75	0.981	-2.28-3.78

Groups	n	Mean height of the germinal epithelium (μm)	SD	95% CI
3 months	360	111.23	20.66	109.09-113.37
6 months	360	108.82	20.99	106.64-110.99
9 months	360	109.12	20.79	106.96-111.27
12 months	360	109.26	22.13	106.97-111.56
18 months	360	100.06*	16.03	98.27-101.86
24 months	360	108.21	12.92	106.87-109.55

Comparison		Difference between the mean heights of the germinal epithelium (μm)	P value	95% CI
Group	Groups
3 months	6 months	2.41	0.545	-1.68-6.51
	9 months	2.11	0.682	-1.98-6.21
	12 months	1.97	0.744	-2.12-6.07
	18 months	11.16	0.000*	6.90-15.43
	24 months	3.02	0.285	-1.07-7.12
6 months	9 months	-0.29	1.000	-4.39-3.80
	12 months	-0.44	1.000	-4.54-3.65
	18 months	8.75	0.000*	4.48-13.01
	24 months	0.60	0.998	-3.48-4.70
9 months	12 months	-0.14	1.000	-4.24-3.95
	18 months	9.05	0.000*	4.78-13.31
	24 months	0.90	0.998	-3.19-5.00
12 months	18 months	9.19	0.000*	4.93-13.46
12 months	24 months	1.05	0.978	-3.04-5.15
18 months	24 months	-8.14	0.000*	-12.40--3.87

Groups	n	Mean (μm² )	SD	95% CI
3 months	360	8540.59	723.42	8465.60-8615.57
6 months	360	8475.71	711.42	8401.97-8549.45
9 months	360	8533.54	688.97	8462.13-8604.95
12 months	360	8457.50	731.20	8381.71-8533.29
18 months	360	6726.21*	825.90	6640.61-6811.82
24 months	360	7395.75*	707.39	7322.43-7469.07

Comparison		Difference between mean tubular areas (µm² )	P value	95% CI
Group	Groups
3 months	6 months	64.87	0.843	-90.89-220.65
	9 months	7.04	1.000	-148.72-162.82
	12 months	83.08	0.650	-72.68-238.86
	18 months	1814.37	0.000*	1658.59-1970.14
	24 months	1144.83	0.000*	989.06-1300.61
6 months	9 months	-57.82	0.898	-213.60-97.94
	12 months	18.20	0.999	-137.56-173.98
	18 months	1749.49	0.000*	1593.72-1905.27
	24 months	1079.96	0.000*	924.18-1235.73
9 months	12 months	76.03	0.732	-79.73-231.81
	18 months	1807.32	0.000*	1651.54-1963.09
	24 months	1137.78	0.000*	982.01-1293.56
12 months	18 months	1731.28	0.000*	1575.51-1887.06
12 months	24 months	1061.75	0.000*	905.97-1217.52
18 months	24 months	-669.53	0.000*	-825.30--513.75

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[1] Correspondence:
Eduardo Pereira de Miranda Rua Juci Cidade, 121/501 88701-425 Tubarão - SC Brasil Tel.: (55 48)99101-0656 eduardo.miranda1982@gmail.com