Kadam, K.M.

Manjula, A.

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Proc. Indian Acad. Sci. (Anim. Sci.), VoL 94, No.4, August 1985, pp. 427--433.© Printed in India.Effect ofcastration and androgen treatment on the androgen dependentparameters in the accessory glands of the slender loris, lorisTardigradus lydekkerianus (Cabra),A MANJULA* and K M KADAMDepartment of Zoology, Bangalore University, Bangalore 560056, India.• Karnataka State Sericulture Development Institute, Thalaghattapura, Bangalore 560062,India.MS received 29 November 1984; revised 26 March 1985Abstract. The accessory glands ofreproduction in slender loris exhibit reduction in secretoryactivity following castration for 15 days and 30 days. Administration of androgens(testosterone-propionate and 511 dihydrotestosterone) to castrated animals had a differen-tial effect in maintaining the biochemical parameters like citric acid and fructose.Keywords. Loris Tardigradus Iydekkerianus; slender loris; castration; androgen treatment.1. IntroductionOf the several secretions of the male sex accessory glands of mammals, fructose andcitric acid are important and are found in large quantities in the semen. Their synthesisand secretion are entirely regulated by the androgens secreted by the testis (Price andWilliam Ashman 1961; Prasad et al 1973a,b). Amongst the non-human primates,besides the monkey (Dinakar et aI1974a,b) not much information about the prosimianprimates is available. The present investigation deals with the secretions and theirandrogen control in the accessory sex glands ofthe male slender loris, Loris tardiqraduslydekkerianus (cabra).2. Materials and MethodsLorises, also known by the native names as ceylon sloths, sherminds, unhappen lumaand thevangu, occur in some of the forested areas of Southern India and Ceylon fromsea level to an elevation of about 1800 inches. Lorises are found mostly in casurina,tamarind and pongamia trees, quite close to rural habitation. They were collected fromthe forests around Bangalore, brought to the laboratory, maintained in cages andprovided with food and water.Adult male lorises used for the experimental groups were divided into nine groups of3/5 animals in each (table 1).They were castrated by opening the inguinal passage underasceptic conditions using sodium pentobarbitone (Nembutol, Abbot laboratories) asanesthetic. Through a small incision in the inguinal passage, the testis were exposed,carefully freed from the epididymis after ligating the efferent ductules and testicularblood vessels without damaging the vascular supply to the epididymis and ductusdeferens. The different doses of testosterone propionate (TP) used were 125 j.lg, 250 j.lg427428 A Manjula and K M KadamTable 1. Changes in the weights (mg) of the accessory glands in the castrated,testosterone propionate (TP), and 5Ot-dihydrotestosterone (5Ot-DHT) treatedlorises (Mean ±S.E.)BodyweightTreatment (g) Seminal vesicles Prostate gland Cowper's glandsIntact control 279(5) 293-4 ±6·3 75·4 ± 14·9 104·0 ± 3·2$ for 15 days 252(3) 195-4 ±9·0c 55·2 ± 7·0< 85·4 ± 12·4<$ for 30 days 260(3) 192·1 ±4·8b 53-3 ± 3'6b 83·9 ± IHb$ + 500 Jlg of TP/day 241(5) 315·0 ± 38·4b 137·0 ± 9-d' 122·7 ± 9·2b$ + 250 Jlg of TP/day 283(5) 393·0 ± 48·0< 71·7 ± 3·5a 174·1 ± 3·l b$ + 125Jlg of TP/day 259(5) 292·2 ± 89·7a 70·7 ± 19·3a 114·5 ± l3-7a$ + 250pg OfDHT/day 285(5) 299·1 ± 25·6a 71·3 ± 10·5a 117·0 ± 18·if$ + 50 Jlg of DHT/day 284(5) 286·0 ± 20·4a 56·5 ± 6·4 116·0 ±21·5a$ + 5 Jlg of DHT/day 272(5) 216·6 ± 10·4 58·9 ± 3·8 74·3 ± 7·9Figures in parentheses represent the number of animals used in each group. Levels ofsignificance compared with intact control animals. "P« 0·05;' P < 0001; "P« 0-05. t/= Castrated.and 500 Jlgjday and of Soc- dihydrotestosterone (Soc-OHT) were 5 ug; 50 Jlg and250Jlgjday. The androgens were administered subcutaneously in (}2 ml of olive oildaily for 15 days from the next day of castration. Intact and castrated lorises ofcomparable body weights received the vehicle only and they served as controls.The lorises were autopsied 24 hr after the last injection; one castrated group withoutandrogen treatment was autopsied 30 days after castration. The accessory glands wereremoved, cleaned of fat and connective tissue and weighed to the nearest 0·1 mg in atorsion balance; fructose was estimated in the accessory glands by the method of Roe(1934)as modified by Linder and Mann (1960)and citric acid by the method ofEttingeret al (1952). The results were analysed statistically using student's t test.3. ResultsResponse ofthe accessory glands ofreproduction in the lorises to the administration ofdifferent dosages ofTPj5oc-oHTin terms ofchange in weight is shown in table 2.Changesin the content and concentration of fructose in the accessory glands resulting fromcastration and androgen replacement are shown in table 3. In loris, fructose is secretedby the prostate gland in maximum quantities. The seminal vesiclesand Cowper's glandsalso secrete some quantities of the fructose. Castration for 15 days resulted insignificant decrease of fructose concentration in the accessory glands and a furtherdecrease was observed in 30-<iay castrated animals. More than 500 Jlg of TP or 5 Jlg ofSoc-oHTjday maintained the fructose concentration in the seminal vesicles but there wasa three-fold increase in the fructose concentration in the animals treated with 50 Jlg of51X-OHT. Cowper's gland required 250 Jlg of TP or more than 50Jlg of SOC-OHT for themaintenance of normal fructose concentration and in the prostate gland, it required250 p.gof TP/5oc-oHTjday.Changes in the levels of citric acid in the accessory glands of loris resulting fromcastration and androgen replacement is shown in table 3. Citric acid is mainly secretedTable2.EffectofadministrationofTPandDHTonthecontentandconcentrationoffructoseintheaccessoryglandsofthecastratedslenderloris(Mean±S_E)SeminalvesiclesProstateglandsCowper'sglandsA8A8A8Intactcontrol199-0±2-869-4±15-6267-9±99-2140-3±51-9132'5±11-779-2±50-6<$forISdays149-6±20'3d42-5±2NdI37-7±1'3d84-0±15-3d94'5±6-6d54-8±6-0d<$for30days34-1±4-4d16-3±12-3d62'2±9·4d18-1±8-oJ5(}6±19-5d3(}8±15-0d<$+500~gofTP/day3000±21'4b131·8±28·8"30209±151-9"126·2±33-2267-6±121-3"66·2±12-7'~+250~gofTP/day241-2±137-ld64-1±14-0"218-2±94·887-5±24-015N±51-6"63-0±15-9~+125~gofTP/day214-8±74-352-9±5-797-1±43-785-3±29-9I08-2±6-652-0±7-9<$+250~gofDHTIday267-2±50'2b179-1±95-0"690-4±318-2"347-9±88-0"282-0±37-2b288-9±68-0"~+50~gofDHT/day213-0±82'4"162-2±59-3"164-5±45-768-5±2-6144-3±25-7"69-9±24-8'~+5~gofDHT/day203-0±16-6"55-7±29-3131-3±470454-8±12-951-6±19.523'8±5-4Levelsofsignificancecomparedwithintactcontrolanimals,QP<(}05,bP<0-01,cP.s;;(}05,dp<0-05A=~gofFructose/organ,8=~gofFructose/IOOmgoftissue.~ ~ ... 0- :::s § ~ l:l S- ci (tl ~ ~ ~ :::s ... 5- '" ~ ~ c ... (;;. t: 10Table3.EffectofadministrationofrsandDHTonthecontentandconcentrationofcitricacidintheaccessoryglandsoftheslenderloris(mean±S.E)SeminalvesiclesProstateglandsCowper'sglandsABABABIntactcontrol501'3±282-3430-1±42'0478·5±136·6375-8±44-4373-6±223'1195'0±32'8$for15days174'6±115·9"129'5±88·0"166'1±20-7"134'9±46·8"133·7±22·7"65'0±0-0"$for30daysAbsentAbsentAbsentAbsentAbsentAbsent$+500IlgofTP/day481·5±39·4'481'9±39·4"131'3±21·9157-8±0-0302'0±58·6135-8±2'1$+250IlgofTP/day231-1±105'1129·8±0-07H±25·0126'2±10-458'6±12·1126'3±0-0$+1251lgofTP/day110-9±51-4118'0±7·046'3±12'592-9±16·042'5±20-215·1±2·0$+250IlgofDHT/day799-6±217'2b143-8±38·0378·1±26·6353'6±23'0225-9±24·9310·4±87·6"$+50llgofDHT/day437-9±264'0134'6±94'9167-9±1·9191'5±9·0101'3±32'1232-4±58'8"el+51lgofDHT/day305-8±55-371'6±43'1166·7±1·5107-8±3H41·8±10-478·6±HLevelsofsignificancecomparedwithintactcontrols"P<0-05,bP<0-01,'P:E;0-05,"P<0'05A=Ilgofcitricacid/organ.B=Ilgofcitricacid/l00mgoftissue.~ tH o ;l... E::: ~. s:: Q ~ ~ E::: tCastration and androgen treatment in slender loris 431by the seminal vesicles in maximum quantities. Like fructose, citric acid in the accessoryglands also depends on the androgens. Prostate and Cowper's glands also contributecitric acid to the seminal plasma in the slender loris. In the IS-day castrated animals theconcentration of citric acid had decreased significantly and it was completely absentfrom the glands of 30-day castrated animals. Requirement of androgens for themaintenance ofcitric acid concentration by the different glands varied. SOO Jig of TP or2S0 Jig of 5a-OHT maintained the citric acid concentration in the seminal vesicles whilemore than SOO Jig of TP or more than 2S0 Jig of Sa-oHT was required for themaintenance of the citric acid concentration in the prostate and Cowper's glands.4. DiscussionA decrease in the weights and secretory activity of the male reproductive system ofslender loris has been observed following castration (Manjula and Kadam 1980) for15 days. These tissues showed a further significant decrease in weight after 30 days ofcastration. A similar decrease has been observed in laboratory rodents (Price andWilliam Ashman 1961; Mann 1964;Gupta et a11974), hamster (Ortiz 19S3;Karkun etal 1974), and rhesus monkey (Dinakar et alI974a,b).Fructose has been found in the semen ofman, monkey, ram, guinea pig, rat and othermammals (Mann 1964). Androgen is necessary for the production of fructose (Mann1964; Gassner and Hopwood 19S2) in the male reproductive system. Fructose issecreted mainly by the prostate gland in the slender loris, but the seminal vesicles andCowper's gland also contribute. In the castrated guinea pig and rats (Harold and Clara1955a,b) a lower dosage of testosterone (Q-SJIg) was sufficient to restore the fructosecontent. In the accessory glands of mouse, fructose level had decreased significantlyfollowing castration for 3 days (Mawhinney et al1970). In the castrated loris, a higherdose ofandrogens, SOO JigofTPand 2S0 JigofSa-oHT, was required by the prostate andCowper's glands to restore the secretory activity in IS-day castrated animals. Theseminal vesicles required 2S0 Jig of TP and a lower dose of Sa-OHT (S Jig). Thus Sa-OHTappears to be more potent than TPin restoring and maintaining the secretory activity ofthe accessory glands. Similarly in the rhesus monkey, the seminal vesicles required fourimplants of Sa-OHT and eight implants of testosterone (Dinakar et aI1974a,b). Thecontent of fructose in dorsolateral prostate was maintained at control levels withSOO JIgofTPor 2S0 JIgofSa-oHTin rats (Guptaet aI1974). In loris, as in monkey and rat,Sa-oHTis more potent, whereas in hamster (Karkun et al1974) testosterone appears tobe more potent. This shows that testosterone and Sa-OHT were not equipotent instimulating the production of fructose in the slender loris.Citric acid is also an androgen-dependent parameter in theaccessory glands. Most ofthe higher mammals (Barron and Huggins 1946a,b; Humphrey and Mann 1948, 1949;Schersten 1929, 1936), have a high concentration of citric acid in semen. A directrelationship exists between the plasma testosterone and citric acid in seminal plasma inman and it is a good index of androgen secreted by the gonads (Dondero et al 1972).Decrease in the levels of citric acid in the seminal vesicles of rabbit followingcastration has been observed (Humphrey and Mann 1948)and it reappears followingadministration of testosterone. In the accessory glands of loris, castration for 15 daysresulted in a significant decrease in the levels ofcitric acid and it was completely absent432 A M anjula and K M Kadamin the seminal vesicles of 30-day castrated animals, prostate and Cowper's glands alsoshowed significant reduction in the citric acid content. In the castrated loris, seminalvesicles required a low dosage of 5ex-DHT (50 Jig) and a high dosage of TP (500 Jig)for themaintenance of citric acid content. The prostate and Cowper's glands required morethan 500 Jig of rz or 250 Jig of 5ex-DHT, of both the androgens, 5ex-DHTis more potent inlow doses than TP in maintaining citric acid content in the seminal vesicles thusindicating greater sensitiveness to low doses of 5ex-DHT, than to testosterone itself.Androstendione and 5fJ-androsten 3-01-17-one have been found in the seminalvesicles after administration of testosterone (Harding and Samules 1962; Kinson 1962;Evalkar et a/1964) and 5ex-androstane 17-01-3-one is found bound to the macromole-cular fraction of the homogenate of the seminal vesiclesin the ventral prostate of the rat(Unjem and Tveter 1969). In the slender loris the results indicate that 5ex-DHT is morepotent than TP and hence it is possible that similar DHT binding sites to macromoleculesare present in the accessory glands of loris also.AcknowledgementOne of the authors (AM) is grateful to the UGC, New Delhi, for the award of afellowship.ReferencesBarron E S G and Huggins C 1946a The citric acid and aconitase content of the prostate; Proc. Soc. Exp. BioI.Med. 63 85-87Barron E S G and Huggins C 1946b The Metabolism of the prostate transamination and citric acid; J. Urol.55385-390Dinakar N, Renu A and Prasad M R N 1974aEffects of microquantities of testosterone on the epididymisand accessory glands of castrated rhesus monkey, Macaca Mulaua; J. Endocrinol. 60 399-408Dinakar N, Arora R and Prasad M R N I974b Effects of micro quantities of 5cx-dihydrotestosterone on theepididymis and accessory glands of the castrated rhesus monkey, Macaca mulat!a; Int. J. Fert, 19133-139Dondero F, Sciarra. F and Isidori A 1972 Evaluation of relationships between plasma testosterone andhuman seminal citric acid; Fen. Ster. 23 168--171 .Ettinger R H, Goldbaum L H and Smith L H 1952 A simplified photometric method for the estimation ofcitric acid in biological fluids; J. Bioi. Chem. 199 531-536Evalker K 0, Mosebach 0 and Dirscherl W 1964 Acta Endocr. Kopen. 45 437Gassner F X and Hopwood M C 1952 Seminal amino acid and carbohydrate pattern of bulls with normaland abnormal testis function; Proc. Soc. Exp. Bioi. Med. 81 37-43Gupta G, Rajalakshmi M and Prasad M R N 1974 Regional differences in androgen threshold of theepididymis of the castrated rat; Steroids 24 575-586Harold A L and Clara M S 1955a The effect ofandrogens on fructose production by the sex accessories ofmale guinea pig and rat; Endocrinology 59 404-412Harold A L and Clara M S 1955b Effects of castration and androgen administration on metaboliccharacteristics of the guinea pig seminal vesicles; Am. J. Physiol. 183 371-376Harding B Wand Samules L T 1962 The uptake and sub cellular distribution of C 14-labelled steroid in ratventral prostrate following in vivo administration of C'<-testosterone; Endocrinology 70109-118Humphrey G F and Mann T 1948 Citric acid in Semen. Nature (London) 161 352-353Humphrey G F and Mann T 1949 Studies in the metabolism of semen: Citric acid in semen; Biochem. J. 4497-105Karkun T B, Rajalakshmi M and Prasad M R N 1974 Maintenance of the epididymis in the castrated goldenhamster by testosterone and dihydrotestosterone; Contraception 9 417Castration and androgen treatment in slender loris 433Kinson G 1962 Fed. Proc. 21 24Linder H R and Mann T 1960 A relationship between the content of androgenic steroids in the testis andsecretory activity of the seminal vesicles of bull; J. Endocrinol. 21 341-360Manjula A and Kadam K M 1980 Effect of testosterone propionate and 5cx-dihydrotestosterone onepididymis and aocessory glands of the castrated slender loris. Loris cardigradus lydekkerianus (cabra);Indian J. Exp. BioI. 18 466-470Mann T 1964 Biochemistry of semen and of the male reproductive trat (London: Methuen)Mawhinney M G, Knytck J RET and Thomas] A 1970 Relationship of fructose and fructose phosphateesters in the aocessory sex organs of the mouse; J. Endocrinol. 46 545-546Ortiz F 1953 The effects of castration on the reproductive system of the golden hamster; Anat. Rec. 11765-93Prasad M R N, Rajalakshmi M, Gupta'G and Karkun T 1973a Control of epididymal function; J. Reprod.Fert. Suppl. 18215-222Prasad M R N, Rajalakshmi M, Gupta G, Dinakar N, Arora A and Karkun T 1973b Epididymalenvironment and maturation of spermatozoa. Presented at the lnt. Conf. for the study of fertility andsterility in the male, Milan, ItalyPrice D and William Ashman H G 1961The aocessory reproductive glands of mammals. In: Sex and internalsecretions eds. (Baltimore: W C Young Williams and Wilkins Co.) Vol. 1, pp. 366, III ed.Roe] H 1934A colorimetric method for the determination of fructose in blood and urine;J. Bioi. Chem. 10715-22Schertsen B Skandinav 1929 Arch. Physiol. 58 90Schertsen B Skandinav 1936 Arch. Physiol. 74 90Unjem 0 and Tveter K U 1969 Preliminary characterisation ofan androgen macromolecular complex fromthe rat ventral prostate; Acta Endocrinol. Copenh. 62 153-154

Effect of testosterone propionate and 5α-dihydrotestosterone on epididymis and accessory glands of the castrated slender loris Loris tradigradus lydekkerianus (Cabra)

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Effect of testosterone propionate and 5α-dihydrotestosterone on epididymis and accessory glands of the castrated slender loris Loris tradigradus lydekkerianus (Cabra)

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