== Ab stract =
Cathepsin D Expression in Intestinal Ganglion Cells of Neonate
Dae-Yeon Kim, M.D., Seong-Cheol Lee, M.D., Kwi-Won Park, M.D., Woo-Ki, Kim, M.D.
Department of Surgery, Seoul National University College of Medicine, Seoul, Korea
Diagnosing Hirschprung's disease is one of the clinical challenges of this disorder. In the stomach and the intestines, Cathepsin D was readily detected in cytoplasm of the rat gastric and in intestinal ganglion cells of the autonomic nervous system. The objectives of the present study were to examine cathepsin D expression in ganglion cells of the submucosal and myenteric plexuses of the intestine of children and to determine the utility of immunohistochemical staining of cathepsin D for detection of immature ganglion cells. Paraffin blocks of 35 intestinal segments were reviewed for immunohistochemical staining with polyclonal antibody to cathepsin D and hematoxy lin-eosin stainings from the compatible specimens. There were 9 aganglionic segments and 9 ganglionic segments of neonates with Hirschsprung's disease, 8 intestinal segments with non-Hirschsprung's disease in neonates and 9 intestinal segments with non-Hirschsprung's disease infants over the age of 10 months. All ganglion cells showed intense granular cytoplasmic reactivity for cathepsin D regardless of maturity and all aganglionic segments had no expression for cathepsin D in the submucosal and myenteric plexuses of the intestine. However, histiocytes within the laminar propria and submucosa stained positively for cathepsin D. In conclusion, intestinal ganglion cells in children have reactivity for cathepsin D, threrfore immuno histochemical staining for cathepsin D can be used for identification of ganglion cells in neonates.
Index Words: Cathepsin D, Hirschsprung's disease, Ganglion cell, Neonate
Corresspondence ; Dae-Yean Kim, M.D., Department oj Surgery, Seoul National University College of Medicine, 28, Yunku/l-dong, Chongro-Ku, Seoul 110-744, Korea
- 39 - 01] 01~ A11r11 i2.. ~~-6}~ ~o] uH- ~.R-6}q9. Cathepsin D~ ~~{} 7Cl111 ~ .¥. W~ ~ 2. 71-¥!- 011 ~A~-6}~ lysosomal aspartic proteinase .£ A}'it -6]-'f-41::::E.;g-~ 2. 1887\1 Hirschsprungo] .!f-{} 01]A1:: E11'\~ 12?lf-E.~ l..}E}l..}71 A 1 .3}~ qlO. Cathe 7Cl~ 71r11~;AJ(aganglionic megacolon)o]2-]-.Jl ;;.~ g psin D~ ~~:3J, ~-6] {}7Cl-'t!(neuron)o1] %If--6}.JL, 1 .2-i2.. ~31~ 0]4- , 1948\1 Whitehouse9} Kernohan {}7Cl~A~1.¥.7} 01 .K~~ ~-61 ~01 .¥.W-6}.JL ~q 2. o]:@.{! ~~lf- ;AJ91 ~~-6}9} ~{}7Cl%01] {}7Cl :: ~2. ~, '7], A}'it91 ~~~:3JJl} '7]91 4-~ {} ~A~].¥.7} 'irt:: {i{iAa ~:@'?:l ~ 'il-r~cf -6].s41:::: 7Cl~(dorsal root ganglion)Jl} ~'To1lA1 '{J~!r.J~.Jl, E.;g- ~.g. 1948\1 SwensonJl} Bill 0] ;;.~ g.2-i2.. ~ "8' '7]91 ;AJ {l7Cl ~ A~].£ o1lA1 cathepsin D 7} ~7t! -6} :: :3J;AJA~{l.2-i2.. ZlB-6}.Jl 7.]liJ,jJ{J ~ 'il-r~ 0]4- ~ ~ 2. 'il-ri>1 A=j ~ qll-l3. 2. ~E11 ,\~c.]~~~ ~~01] ~~{J~ '?:lAJ~~ ~ A~ :A}%.g. cathepsin D 7} ~ o} 91 ;AJ {} 7Cl ;
Table 1. Materials and Tissue with Ganglion Cells Immunoreactive to Cathepsin D No. of No_ of posi Tissues Age range Site cases tive reaction Mature ganglionic segment imperforate anus 9 10-17m 2 ileum. 7 colon 9 Ganglion segment in neonate necrotizing enterocolitis 4 2- 28d All ileum 4 ileal atresia 4 1- 2d All ileum 4 Hirschsprung'disease 9 2-28d 1 jejunum. 1 ileum. 7 colon 9 Aganglionic segment in neonate Hirschsprung'disease 9 4- 28d 2 ileum. 2 appendix. 5 colon o
- 40- Jl, ;<.J- ul *iJ:2. ll~~ g .!i!.01t?1 {l7cl1H~~ Jiil~ t>}'j q12,13(.::I ~ 3, 4).
1. Hematoxylin-eosin ~ ~.£.£ {l7cl ~Ail ~ ~ ~ '?H!- 'T- 9,.l, ~ g ~-3j 2. cathepsin D t?1 ~ ~-3j §} '§{ ~~OJI 1fr g t>}'jqClE 1). 2. "'a-Si- {l7cl~Ail~7} ul"'a-Si- {l7cl~Ail~.!i!.q Ail~ ~o l %lJLt>}'jJl "ilAil~ 3l}~~91 'i:l~~Cch romatin)3l} % [:-1 %.!i!. °]711 %{l OJ] ~.7.1 ~ ~ ~?ll Fig. 1. Mature colonic ganglion cells in 17 month-old 5'.. T~O] ~ ~~?1°J: .s:.~ cathepsin DOJl 7J-~ male(H & E, X 1,000). t?1~ 1frg g .!Jl..'jqC.::I~ 3,4). 3. Y1 IlHl~, "iJ:s:I}.:r-:::: cathepsin DOJI t?1 ~ 1fr g
Fig. 2. Cathepsin D immunohistochemical staltung of mature colonic ganglion cells in 17 month-old male ( x 1000). This figure shows deep yellow granules Fig. 3. Immature intestinal ganglion cells in 2 day crowded the cytoplasm. -old female(H & E, x 1,000).
~-3j~ -'t!.£.£ hematoxylin-eosin ~~ ~ A]i;~t>}Jl %A] OJ] ~,;;,}tiJ-~llCactive cathepsin D isolated from human liver)9} o],;;,}tiJ-?l]CDAKO poly clonal rabbit anti-cathepsin D)~ ;+%t>}9 cathepsin DOJ] r11 ~ t?1 ~ ~-3j §}'§{~ ~ ~ Al i;~t>}'j It.
1) Hematoxylin-eosin ~~ :C{l7cl ~ 3l} 13 ~t>}%OJl Ai Ail ~~ °1 %lJL ~ qzj ~.£.£ %-2- -'t!1P'a ~:i!} !f~~ ~:±'~11% 7}?IJl 9,H::: Ai] ~ ~ {l7cl1~A ~£ :sa~ t>}'j q\::l ~ 1, 2). Fig. 4. Cathepsin D immunohistochemical staining of immature intestinal ganglion ceJls in 2 day-old female 2) Cathepsin DOli (He!- ~~~~£:I-~~~ (X 1,000). This figure shows granular localization of Ai]~~ol 3l}~~91 ~~~.£.£ zlA711 ~~~l cathepsin D in the perinuclear cytolasm and rosette -like arrangement.
- 41 - "8"};<'] U~q. q21 4. Jl %~J!j- 1:1 ~"8"} '(toJ] ~ ~ 3:31 T~ cath "8"] .s.~~n:~~ oJl Ai AJ£1 ~~~. {l75 ;<']IlH ~ ~ epsin DoJ] ~~ ~.%% ~o ] ;<.]'?l-, 'i±~ T3:°]Jl ~oa"8"} Jl 7-J {l75-2-Acj <2:jAl]~ ~~ o] 7]~~ ~~ ~75~A~]~oJ]Ai ~o] ~ J!j-ifl t§.91 A~] ~~~ 'Zl ~~ 0] q. ; - 42 - ~9j A11.¥.~~~ ~7cl ~ iiiJl, ~AH:!- ~~~31} -e Surg 202:349-355, 1985 ~{JAd ~~711 ~ ;1-;7,1 \1 0} {l7cPJA11.¥..2t T-~01 7. Dahams BB: The gastrointestinal tract, in Stoc 01 ~ 7-1 ~l9)-ct . ker JT, Debner LP: Pediatric Pathology. Lipp Cathepsin D 7 I- ~~{l ~ 111 ~ .A}%{l ~ 1119-] "'d incott, Philadelphia, PA, 1991, P 670 Si-31}~ol ] o~7H-oI--c ~~.£ ~?jA4 ~J-c [11, ~~.£ 8. Venugopal S, Mancer K, Shandling B: the va -c ::J. 3ll-~ol l }.i 9-1 ~~~ 7 ] *~ ~~3lj- ll"'J ~ lidity of rectal biopsy in relation to morpho - 43- - .l:o}9.Ji!J- ~]5 -t! ;<1]1 I 1999';1 - sease. J Pediatr Surg 7: 11-17, 1972 20. Tam PK: An immunochemical study with ne 17. lkawa H, Kim SH, Hendren WH, Donahoe uron-specific-enolase and substance P of human PK: Acetylcholinesterase and manometry in the enteric innervation -the normal developmental diagnosis of the constipated child. Arch Surg pattern and abnormal deviations in Hirsch 121:435-438, 1986 sprung's disease and pyloric stenosis. I Pediatr 18. Athow AC, Filipe MI, Drake DP: Problems Surg 21(3):227-232, 1986 and advantages of acetylcholinesterase histoche 21. Larsson LT: Hirschsprung's disease-immunohi mistry of rectal suction biopsies in the di stochemical findings. Histol Histopathol 9(3): agnosis of Hirschsprung's disease. J Pediatr 615-629, 1994 Surg 25:520-526, 1990 22. Yonezawa S, Takahashi T, Wang XI, Wong 19. Robey SS, Kuhajda FP, Yardley JH: Imm RN, Hartsuck lA, Tang I: Structures at the unoperoxidase stains of ganglion cells and ab proteolytic processing region of cathepsin D. I normal mucosal nerve proliferations in Hirsch BioL Chern 263:16504-16511, 1988 sprung's disease. Hum Pathol 19:432-437, 1988 - 44-