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Barr bodies in testis with Klinefelter syndrome
BARR BODIES KLINEFELTER
IN TESTIS WITH SYNDROME
A. K. M. SHAMSUDDIN, CHIK-KWUN
TANG,
M.D.,
PH.D.
M.B.
From the Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland
ABSTRACT - Barr bodies are described in the interstitial cells of testes of a chromatin-positive patient with Klinefelter syndrome. Ultrastructural studies confirm the origin of the Barr body from the nuclear chromatin. Ultrastructurally the interstitial cells showed diminished, smooth endoplasmic reticulum and lipid droplets probably indicating impaired function.
In the evaluation of patients with Klinefelter syndrome buccal mucosa is the usual tissue to look for Barr bodies. Although Ferguson-Smith et al.’ separated the histologic features of the testes in Klinefelter syndrome into chromatinpositive and chromatin-negative groups, testicular Barr bodies were not included in the description distinguishing one group from the other. We have recently encountered nuclear structures that are identical to Barr bodies in the interstitial cells of testes removed from a patient with Klinefelter syndrome and wish to report the histologic and ultrastructural findings.
On gross examination the testes weighed 5.3 Gm. each. They were small and shrunken with Microscopically, the testes thick capsule. showed very thick tunica albuginea. Most of the seminiferous tubules were hyalinized. The re-
maining tubules had thick basement membranes and were lined by Sertoli cells. Only a few seminiferous tubules showed some germ cells. The tubules were surrounded by fibrous tissue in which islands of interstitial cells (Leydig cells) were present (Fig. 1). These interstitial cells were pleomorphic but invariably contained eosinophilic cytoplasm. No definite crystals of Reinke were observed. Prominent nucleoli were seen. In some nuclei, triangular or hemioval basophilic bodies identical to Barr bodies were seen blending with the nuclear membranes. Tissues from one of the two testes for electron microscopic examination were fixed adequately in formaldehyde (4%) and glutaraldehyde (1%) mixture at 176 mOsm., were washed with sucrose cacodylate buffer, osmicated, stained en bloc in uranyl acetate, dehydrated in graded ethanol, and embedded in Epon-812. Onemicron thick sections were stained with toluidine blue and representative areas were selected for thin sectioning. 600-800 A thin sections were stained with lead citrate and uranyl acetate and examined with a JEOL 100 B electron microscope at 60 Kv. Electron microscopic study revealed large interstitial cells with prominent profiles of smooth endoplasmic reticulum some of which were dilated and many others were vesicular. There were scanty mitochondria, some with tubular cristae. Abundant Golgi profiles and stacks of rough endoplasmic reticulum were also
74
UROLOGY
Case Report A thirty-four-year-old Caucasian man was diagnosed to have Klinefelter syndrome with chromosomal studies showing a XXY pattern. He had had retractile testes which caused severe pain when they were retracted and when he tried to lift heavy objects. The patient’s body height was greater than the arm span. He had a normal male hair pattern. The patient was on Depo-testosterone therapy. The testes were removed to relieve his pain. Pathologic findings
/ JANUARY 1980 / VOLUME
XV, NUMBER
1
Section oj FIGURE 1. testis showing hyalinized tubules seminiferous lined by Sertoli cells and groups of interstitial cells with pleomorphic bizarre nuclei (hematoxylin-eosin stain). Inset shows Barr paraffinbody in embedded hematoxylineosin section. (Original magnilfications, x 80, x 1,260, respectively.)
seen, The cc>11membrane showed some microvillous projections (Fig. 2A). Crystals of Reinke were not observed. Triangular clumps of electron-dense granular structures corresponding to the Barr bodies seen on light microscopy were found along the nuclear membrane (Fig. 2B). The granular pattern of these structures was indistinguishable from that of the adjacent chromatin and was distinctly different from that of the nucleoli which were prominent. Lipid droplets were not observed. Abundant finely granular and fibrillar moderately electrondense substance was present in between organelles. Sertoli cells lining the tubules were elongated with relatively straight lateral cell membrane without folding. Desmosomes were conspicuous. There were scanty amounts of organelles, including a few stacks of rough endoplasmic reticulum, irregular lipid inclusions, a few mitochondria, bundles of cytoplasmic filaments and crystalloid of Charcot-Bottcher; the basal lamina appeared to be reduplicated (Fig. 3). Comment
FIGURE 2. (A) Interstitial cell of ikydig showing microvillous projection of cell membrane, stacks of rough endoplasmic reticulum, scanty smooth endoplasmic reticulum, no lipid, and tubular cristae of mitochondria. (B) Nucleus of interstitial cell showing trapezoid area of clumped chromatin (left side); chromatin material considered to be Barr body has morphology identical to rest of chromatin. (Original mugnifiations X 4,000 and X 8,000 respectively.)
UROLOGY
/ JANUARY 1980 / VOLUME
XV, NUMBER
1
As described by Ferguson-Smith et al., 1 the testes from chromatin-positive cases showed aggregates or masses of interstitial cells with pleomorphic nuclei. Large areas devoid of tubules were found. The remaining tubules were either totally hyalinized or lined by Sertoli cells. Spermatogenesis was rare. The testes from the chromatin-negative cases showed diffusely increased interstitial cells that were cytologically normal. The seminiferous tubules were of normal size, but most were lined solely by Sertoli cells. In all cases, a few tubules showed some spermatogenesis. The
75
FIGURE 3. Seminiferous tubule showing reduplicated basal lamina, tall elongated Sertoli cells, lipid vacuole, stacks of rough endoplasmic reticulum, straight lateral membrane, cytoplasmic filaments, and Charcot-Biittcher crystalloid (original magnijcation x 4,000). Inset: Higher magnijcation of Charcot-B6ttcher crystalloid.
light microscopic examination were indistinguishable from that of the adjacent chromatin. This finding, in conjunction with an extra number of sex chromosomes, indicates that Barr bodies are indeed sex chromosomes. The function of interstitial cells is known to be impaired to varying degrees despite the fact that they are hyperplastic.2 In our patient, the impaired function of the interstitial cells is suggested by the ultrastructural findings when compared with those of interstitial cells in a man of normal androgenic status.3 Although profiles of smooth endoplasmic reticulum were prominent in the interstitial cells of the testis under study, they were not as packed as those in interstitial cells that actively produced androgenie hormone. The reduction of smooth endoplasmic reticulum, accompanied by the lack of lipid droplets which are conspicuous in active interstitial cells suggests that this patient’s interstitial cells probably were not as active as normal interstitial cells. However, due to lack of correlative laboratory data, no valid conclusion can be drawn on morphologic changes alone. Whether or not the presence of the finely fibrillar material in between the organelles reflects an impaired function of the interstitial cells is not known. As in normal Sertoli cells, the Sertoli cells in this testis contained lipid droplets in cytoplasm, bundles of cytoplasmic filaments, desmosomes, and Charcot-Biittcher crystalloid.4 Different from the normal Sertoli cells, however, were scanty, smooth endoplasmic reticulum and stacks of rough endoplasmic reticulum. Lamellar bodies which are also features of normal Sertoli cells were not found in this testis. Whether or not this morphologic difference reflects functional disturbance cannot be evaluated since the function of the Sertoli cell is not clear. 22 South Green Street Baltimore, Maryland 21201 (DR. SHAMSUDDIN)
light microscopic findings in our patient, therefore, belong to the chromatin-positive category. Additionally, triangular or hemi-oval dark purple structures were found attached to the nuclear membrane of the nuclei of some interstitial cells. These structures demonstrated the same staining characteristics as that of the nucleoli found within the same nuclei, appearing dark purple on hematoxylin-eosin stain, and are interpreted as identical to Barr bodies found in the buccal mucosa. The granular pattern found ultrastructurally in the area corresponding to Barr bodies on
1. Ferguson-Smith MA, Lennox B, Mack WS, and Stewart JSS: Klinefelter’s syndrome. Frequency and testicular morphology in relation to nuclear sex. Lancet 2: 167 (1957). 2. Wang T-W, Straus FH, and Warner NE: Testicular biopsy in the study of male infertility. I. Testicular causes of infertility, Arch. Pathol. 95: 151 (1973). 3. DeKrester DM: The fine structure of the testicular interstitial cells in men of normal androgenic status, Z. Zellforsch. Mikrosk. Anat. 80: 594 (1967). 4. Nagano T: Some observations on the fine structure of the Sertoli cell in the human testis, ibid. 73: 89 (1966).
76
UROLOGY
References
/
JANUARY 1966
/
VOLUME XV, NUMBER
1
IN TESTIS WITH SYNDROME
A. K. M. SHAMSUDDIN, CHIK-KWUN
TANG,
M.D.,
PH.D.
M.B.
From the Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland
ABSTRACT - Barr bodies are described in the interstitial cells of testes of a chromatin-positive patient with Klinefelter syndrome. Ultrastructural studies confirm the origin of the Barr body from the nuclear chromatin. Ultrastructurally the interstitial cells showed diminished, smooth endoplasmic reticulum and lipid droplets probably indicating impaired function.
In the evaluation of patients with Klinefelter syndrome buccal mucosa is the usual tissue to look for Barr bodies. Although Ferguson-Smith et al.’ separated the histologic features of the testes in Klinefelter syndrome into chromatinpositive and chromatin-negative groups, testicular Barr bodies were not included in the description distinguishing one group from the other. We have recently encountered nuclear structures that are identical to Barr bodies in the interstitial cells of testes removed from a patient with Klinefelter syndrome and wish to report the histologic and ultrastructural findings.
On gross examination the testes weighed 5.3 Gm. each. They were small and shrunken with Microscopically, the testes thick capsule. showed very thick tunica albuginea. Most of the seminiferous tubules were hyalinized. The re-
maining tubules had thick basement membranes and were lined by Sertoli cells. Only a few seminiferous tubules showed some germ cells. The tubules were surrounded by fibrous tissue in which islands of interstitial cells (Leydig cells) were present (Fig. 1). These interstitial cells were pleomorphic but invariably contained eosinophilic cytoplasm. No definite crystals of Reinke were observed. Prominent nucleoli were seen. In some nuclei, triangular or hemioval basophilic bodies identical to Barr bodies were seen blending with the nuclear membranes. Tissues from one of the two testes for electron microscopic examination were fixed adequately in formaldehyde (4%) and glutaraldehyde (1%) mixture at 176 mOsm., were washed with sucrose cacodylate buffer, osmicated, stained en bloc in uranyl acetate, dehydrated in graded ethanol, and embedded in Epon-812. Onemicron thick sections were stained with toluidine blue and representative areas were selected for thin sectioning. 600-800 A thin sections were stained with lead citrate and uranyl acetate and examined with a JEOL 100 B electron microscope at 60 Kv. Electron microscopic study revealed large interstitial cells with prominent profiles of smooth endoplasmic reticulum some of which were dilated and many others were vesicular. There were scanty mitochondria, some with tubular cristae. Abundant Golgi profiles and stacks of rough endoplasmic reticulum were also
74
UROLOGY
Case Report A thirty-four-year-old Caucasian man was diagnosed to have Klinefelter syndrome with chromosomal studies showing a XXY pattern. He had had retractile testes which caused severe pain when they were retracted and when he tried to lift heavy objects. The patient’s body height was greater than the arm span. He had a normal male hair pattern. The patient was on Depo-testosterone therapy. The testes were removed to relieve his pain. Pathologic findings
/ JANUARY 1980 / VOLUME
XV, NUMBER
1
Section oj FIGURE 1. testis showing hyalinized tubules seminiferous lined by Sertoli cells and groups of interstitial cells with pleomorphic bizarre nuclei (hematoxylin-eosin stain). Inset shows Barr paraffinbody in embedded hematoxylineosin section. (Original magnilfications, x 80, x 1,260, respectively.)
seen, The cc>11membrane showed some microvillous projections (Fig. 2A). Crystals of Reinke were not observed. Triangular clumps of electron-dense granular structures corresponding to the Barr bodies seen on light microscopy were found along the nuclear membrane (Fig. 2B). The granular pattern of these structures was indistinguishable from that of the adjacent chromatin and was distinctly different from that of the nucleoli which were prominent. Lipid droplets were not observed. Abundant finely granular and fibrillar moderately electrondense substance was present in between organelles. Sertoli cells lining the tubules were elongated with relatively straight lateral cell membrane without folding. Desmosomes were conspicuous. There were scanty amounts of organelles, including a few stacks of rough endoplasmic reticulum, irregular lipid inclusions, a few mitochondria, bundles of cytoplasmic filaments and crystalloid of Charcot-Bottcher; the basal lamina appeared to be reduplicated (Fig. 3). Comment
FIGURE 2. (A) Interstitial cell of ikydig showing microvillous projection of cell membrane, stacks of rough endoplasmic reticulum, scanty smooth endoplasmic reticulum, no lipid, and tubular cristae of mitochondria. (B) Nucleus of interstitial cell showing trapezoid area of clumped chromatin (left side); chromatin material considered to be Barr body has morphology identical to rest of chromatin. (Original mugnifiations X 4,000 and X 8,000 respectively.)
UROLOGY
/ JANUARY 1980 / VOLUME
XV, NUMBER
1
As described by Ferguson-Smith et al., 1 the testes from chromatin-positive cases showed aggregates or masses of interstitial cells with pleomorphic nuclei. Large areas devoid of tubules were found. The remaining tubules were either totally hyalinized or lined by Sertoli cells. Spermatogenesis was rare. The testes from the chromatin-negative cases showed diffusely increased interstitial cells that were cytologically normal. The seminiferous tubules were of normal size, but most were lined solely by Sertoli cells. In all cases, a few tubules showed some spermatogenesis. The
75
FIGURE 3. Seminiferous tubule showing reduplicated basal lamina, tall elongated Sertoli cells, lipid vacuole, stacks of rough endoplasmic reticulum, straight lateral membrane, cytoplasmic filaments, and Charcot-Biittcher crystalloid (original magnijcation x 4,000). Inset: Higher magnijcation of Charcot-B6ttcher crystalloid.
light microscopic examination were indistinguishable from that of the adjacent chromatin. This finding, in conjunction with an extra number of sex chromosomes, indicates that Barr bodies are indeed sex chromosomes. The function of interstitial cells is known to be impaired to varying degrees despite the fact that they are hyperplastic.2 In our patient, the impaired function of the interstitial cells is suggested by the ultrastructural findings when compared with those of interstitial cells in a man of normal androgenic status.3 Although profiles of smooth endoplasmic reticulum were prominent in the interstitial cells of the testis under study, they were not as packed as those in interstitial cells that actively produced androgenie hormone. The reduction of smooth endoplasmic reticulum, accompanied by the lack of lipid droplets which are conspicuous in active interstitial cells suggests that this patient’s interstitial cells probably were not as active as normal interstitial cells. However, due to lack of correlative laboratory data, no valid conclusion can be drawn on morphologic changes alone. Whether or not the presence of the finely fibrillar material in between the organelles reflects an impaired function of the interstitial cells is not known. As in normal Sertoli cells, the Sertoli cells in this testis contained lipid droplets in cytoplasm, bundles of cytoplasmic filaments, desmosomes, and Charcot-Biittcher crystalloid.4 Different from the normal Sertoli cells, however, were scanty, smooth endoplasmic reticulum and stacks of rough endoplasmic reticulum. Lamellar bodies which are also features of normal Sertoli cells were not found in this testis. Whether or not this morphologic difference reflects functional disturbance cannot be evaluated since the function of the Sertoli cell is not clear. 22 South Green Street Baltimore, Maryland 21201 (DR. SHAMSUDDIN)
light microscopic findings in our patient, therefore, belong to the chromatin-positive category. Additionally, triangular or hemi-oval dark purple structures were found attached to the nuclear membrane of the nuclei of some interstitial cells. These structures demonstrated the same staining characteristics as that of the nucleoli found within the same nuclei, appearing dark purple on hematoxylin-eosin stain, and are interpreted as identical to Barr bodies found in the buccal mucosa. The granular pattern found ultrastructurally in the area corresponding to Barr bodies on
1. Ferguson-Smith MA, Lennox B, Mack WS, and Stewart JSS: Klinefelter’s syndrome. Frequency and testicular morphology in relation to nuclear sex. Lancet 2: 167 (1957). 2. Wang T-W, Straus FH, and Warner NE: Testicular biopsy in the study of male infertility. I. Testicular causes of infertility, Arch. Pathol. 95: 151 (1973). 3. DeKrester DM: The fine structure of the testicular interstitial cells in men of normal androgenic status, Z. Zellforsch. Mikrosk. Anat. 80: 594 (1967). 4. Nagano T: Some observations on the fine structure of the Sertoli cell in the human testis, ibid. 73: 89 (1966).
76
UROLOGY
References
/
JANUARY 1966
/
VOLUME XV, NUMBER
1